1. Stem Cells and Regenerative Medicine

Activation of the EGFR/MAPK pathway drives transdifferentiation of quiescent niche cells to stem cells in the Drosophila testis niche

  1. Leah J Greenspan
  2. Margaret de Cuevas  Is a corresponding author
  3. Kathy H Le
  4. Jennifer M Viveiros
  5. Erika L Matunis  Is a corresponding author
  1. Department of Cell Biology, Johns Hopkins University School of Medicine, United States
  2. Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, United States
  3. Graduate Program in Biophysics, Stanford University, United States
https://doi.org/10.7554/eLife.70810
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Cite this article
  1. Leah J Greenspan
  2. Margaret de Cuevas
  3. Kathy H Le
  4. Jennifer M Viveiros
  5. Erika L Matunis
(2022)
Activation of the EGFR/MAPK pathway drives transdifferentiation of quiescent niche cells to stem cells in the Drosophila testis niche
eLife 11:e70810.
https://doi.org/10.7554/eLife.70810
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5 figures, 8 tables and 1 additional file

Figures

Figure 1
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Activation of EGFR signaling in adult hub cells causes them to re-enter the cell cycle.

(A) Schematic of the Drosophila testis stem cell niche. Somatic hub cells (green) secrete signals to adjacent germline stem cells (GSCs, orange) and somatic cyst stem cells (CySCs, dark blue). Both types of stem cells divide asymmetrically to produce differentiating daughter cells. Somatic cyst cells (light blue) envelop clusters of spermatogonia (orange), and together they move away from the testis apex as they differentiate. (B) Schematic of the screen for signals that can trigger hub cells to re-enter the cell cycle. Candidate signaling pathway genes were conditionally mis-expressed in adult hub cells using the hub specific driver E132-Gal4, and mitotic hub cells were identified by immunostaining for phospho-histone H3 (PH3) (see main text for details). (C–H) Single confocal sections through the apex of testes immunostained for Fas3 (hub cell membranes, green), PH3 (mitotic chromosomes, nuclear green), and Vasa (germ cells, magenta). Hubs are outlined in white. Mitotic hub cells (white arrowheads) are found in positive control testes (with Rbf knockdown in the hub, C) and in testes over-expressing components of the canonical EGFR/MAPK pathway in the hub (E–H) but not in negative control testes (D). Mitotic cells are also found outside the hub in both control and experimental testes, as expected (yellow arrowheads). Scale bar (in H, for all panels) is 10 μm.

Figure 2
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Activation of EGFR signaling in adult hub cells causes them to convert to CySCs.

(A–D) Single confocal sections through the apex of testes after 8 days of G-TRACE lineage tracing system expression in adult hub cells. Testes were immunostained for RFP (red fluorescent protein, marking current expression of the hub-specific driver E132-Gal4) and GFP (green fluorescent protein, marking both current and past expression of E132-Gal4) and counterstained with DAPI (blue; marks all nuclei). Hubs are outlined in white. (A'-D') Red channel alone, in white; (A"-D") green channel alone, in white. In control testes (A), hub cells expressing the G-TRACE system alone are marked with RFP and GFP, but no cells outside the hub are marked. In testes over-expressing components of the EGFR signaling pathway in the hub together with G-TRACE (B–D), hub cells are marked with RFP and GFP, and cells outside the hub are also marked, either with GFP and low levels of RFP (yellow arrowheads) or with GFP only (white arrowheads). The marked cells outside the hub appear to be cyst lineage cells that arose by conversion of hub cells to CySCs, which no longer express RFP once they lose their hub cell fate. Scale bar in D, for all panels, is 10 μm.

Figure 3
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Negative regulators of EGFR signaling maintain hub cell quiescence and identity.

(A–B, D) Single confocal sections through the apex of testes immunostained for Fas3 (hub cell membranes, green), PH3 (mitotic chromosomes, nuclear green), and either (A–B) Vasa (germ cells, magenta) or (D) Traffic jam (Tj; somatic cell nuclei, magenta). Hubs are outlined in white. Mitotic hub cells (white arrowheads) are found after knockdown of Sprouty (A) or PTEN (B) in the hub or after knockdown of Argos (D) in cyst lineage cells. Mitotic cells outside the hub are also found (yellow arrowhead). (C) Quantification of dpERK levels in the hub in control, Sprouty knockdown, or Pten knockdown in the hub. dpErk levels in the hub are significantly higher when either Sprouty or Pten are knocked down in the hub than in control testes suggesting these proteins normally inhibit MAPK signaling in the hub. A.U., arbitrary units. Black bars indicate the mean and standard error. Unpaired t test, **p < 0.01, ****p < 0.0001. (E–F) Single confocal sections through the apex of testes after 8 days of G-TRACE lineage tracing system expression in adult hub cells. Testes were immunostained for RFP (red, current expression of the hub-specific driver E132-Gal4) and GFP (green, current and past expression of the driver) and counterstained with DAPI (blue; marks all nuclei). Hubs are outlined in white. (E'-F') Red channel alone, in white; (E"-F") green channel alone, in white. After knockdown of Sprouty (E) or PTEN (F) in the hub together with expression of G-TRACE, hub cells are marked with RFP and GFP, and cells outside the hub are also marked, either with GFP and low levels of RFP (yellow arrowheads) or with GFP only (white arrowheads), suggesting that marked cyst lineage cells arose by conversion of hub cells to CySCs. Scale bars in D (for A-B, D) and in F (for E-F) are 10 μm.

Figure 4
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EGFR signaling is important for testis recovery from CySC ablation.

(A–B) Single confocal sections through the apex of testes immunostained for Fas3 (hub cell membranes, green), dpERK (EGFR pathway activation, red), and counterstained with DAPI (nuclei, blue). Hubs are outlined in white. Insets show the red channel alone, in white and enlarged. In control C587-Gal4, Gal80ts testes (A), dpERK levels are high in cyst lineage cells (indicating high levels of EGFR pathway activation) but low in hub cells. In C587-Gal4, Gal80ts > UAS grim testes (B), 2 days after genetic ablation of all CySCs and early cyst cells, dpERK levels are high in hub cells. Scale bar in B (for A-B) is 20 μm. (C) Quantification of dpERK levels in the hub in control (A) and ablated (B) testes. dpErk levels in the hub are significantly higher in ablated testes than in control testes. A.U., arbitrary units. Black bars indicate the mean and standard error. Unpaired t test, **p < 0.01. (D) Bar graph showing the distribution of testis phenotypes in control C587-Gal4, Gal80ts > UAS grim, Egfr+/+ flies ("control Egfr+/+ ablation") and in C587-Gal4, Gal80ts > UAS grim, Egfr-/+ flies ("EGFR-/+ ablation") at 0, 7, or 14 days after genetic ablation of CySCs and early cyst cells. After ablation (0 days), in both control and Egfr-/+ flies, most testes lack all CySCs and early cyst cells but retain a hub and germ cells (GC) as expected (white bars). At 7 and 14 days after ablation, fewer testes have regained CySCs and early cyst cells (black bars) in Egfr-/+ flies than in control flies and there is a significant difference in phenotype distribution. Chi square test, ***p < 0.001, ****p < 0.0001. (E–I) Single confocal sections through the apex of testes at 7 days after ablation, immunostained for Vasa (germ cells, red), Fas3 (hub cell membranes, green), and Tj (somatic cell nuclei, white), and counterstained with DAPI (nuclei, blue), to illustrate the phenotypes listed in (D). Testes that recover CySCs and early cyst lineage cells have Tj-positive nuclei outside the hub (yellow arrowheads); most also contain germ cells (E) but a few contain just a hub and cyst cells (G). Testes that fail to recover CySCs and early cyst lineage cells can retain a hub and germ cells (F) or just a hub (H) or no hub or germ cells (I). Hubs are outlined in white. Scale bar in I (for E-I) is 20 μm.

Figure 5
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Model for cyst lineage recovery after ablation.

(A) In wild type testes, EGF ligands (red circles) are secreted by germ cells and received by cyst lineage cells. The EGFR pathway is repressed in hub cells by the secreted inhibitor Argos (yellow squares), which sequesters EGF ligands, and by intrinsic pathway inhibitors (Sprouty and PTEN). Hub quiescence and identity are maintained. (B) After genetic ablation of all CySCs and early cyst lineage cells, EGF ligands are no longer sequestered by Argos and are received by the hub. The EGFR pathway is activated in hub cells, driving expression of the downstream transcription factor Pointed and its target genes, resulting in hub cell proliferation and conversion to CySCs. As new CySCs are generated, Argos is once again expressed, down-regulating EGFR signaling in the hub.

Tables

Table 1
Hub cell proliferation after EGFR pathway activation.
Gal4 driverUAS line (BDSC #)Days at 31 °C% Testes with PH3-positive hub cells
Controls
E132-Gal4, Gal80tsUAS-Rbf-RNAi (41863)731% (n = 34/108)****
UAS-Rbf-RNAi (36744)729% (n = 53/183)****
UAS-GFP-RNAi (9330 or 9331)70% (n = 0/237)
C587-Gal4, Gal80tsUAS-GFP-RNAi (9331)70% (n = 0/294)
Downstream Effectors and Transcription Factors
E132-Gal4, Gal80tsUAS-Pointed.P1 (869)77% (n = 4/55)**
UAS-Pointed.P2 (399)76% (n = 6/100)***
UAS-Ras85D.V12 (4847)311% (n = 2/18)**
56% (n = 2/36)*
UAS-Rolled (59006)70% (n = 0/92)ns
Receptors
E132-Gal4, Gal80tsUAS-Egfr Type I (9534)71% (n = 1/143)ns
UAS-Egfr Type II (9533)74% (n = 5/117)**
UAS-Egfr λ (59843)7< 1% (n = 1/163)ns
UAS-PvR λ (58496)71% (n = 1/106)ns
UAS-PvR λ (58428)70% (n = 0/156)ns
UAS-InR (8250)70% (n = 0/48)ns
UAS-InR (8263)70% (n = 0/41)ns
UAS-Heartless λ (5367)72% (n = 3/181)ns
UAS-Breathless λ (29045)70% (n = 0/90)ns
Negative Regulators
E132-Gal4, Gal80tsUAS-Sprouty-RNAi (36709)73% (n = 4/144)*
UAS-Pten-RNAi (33643)73% (n = 4/125)*
UAS-Argos-RNAi (28383)7–8< 1% (n = 1/170)ns
C587-Gal4, Gal80tsUAS-Argos-RNAi (28383)7–82% (n = 6/348)*

  1. Percentages in bold are significant compared to the negative control (UAS-GFP-RNAi driven by the same Gal4 driver).

  2. Fisher’s Exact Test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = not significant.

Table 2
Hub cell proliferation upon EGFR and Notch signaling changes.
UAS lines (with E132-Gal4, Gal80ts)SourceDays at 31 °C% Testes with PH3-positive hub cells
Control
UAS-GFP-RNAiBDSC 9330 or 933170% (n = 0/237)*
Egfr knockdown
UAS-Egfr-DNBDSC 536470% (n = 0/103)†,ns
UAS-Egfr-RNAiBDSC 367700% (n = 0/45)†,ns
UAS-Egfr-RNAiBDSC 600120% (n = 0/46)†,ns
UAS-Egfr-RNAiVDRC 432670% (n = 0/29)†,ns
UAS-Egfr-RNAi✝✝VDRC 1071300% (n = 0/43)†,ns
Notch over-expression or knockdown
UAS-N-CABDSC 5200870% (n = 0/48)†,,ns
UAS-N-DNBDSC 516670% (n = 0/54)†,ns
UAS-N-RNAiBDSC 336110% (n = 0/119)†,ns
UAS-N-RNAiBDSC 336160% (n = 0/63)†,ns
UAS-N-RNAiBDSC 352130% (n = 0/29)†,ns
UAS-N-RNAiBDSC 356400% (n = 0/26)†,ns
Egfr over-expression and Notch combinations
UAS-Egfr Type IIBDSC 953374% (n = 5/117)*
UAS-EGFR Type I; UAS-EGFR Type IIBDSC 9533 + 95347–84% (n = 11/301)‡,ns
UAS-N-DN; UAS-EGFR Type IIBDSC 9533 + 516673% (n = 4/138)‡,ns; §,ns
UAS-N-CA; UAS-EGFR Type IIBDSC 9533 + 520086% (n = 12/205)‡,ns; §,ns
Sprouty knockdown and Notch combinations
UAS-Sprouty-RNAiBDSC 3670973% (n = 4/144)*
UAS-EGFR Type I; UAS-Sprouty-RNAiBDSC 36709 + 95347–83% (n = 3/101),ns
UAS-N-DN; UAS-Sprouty-RNAiBDSC 36709 + 516671% (n = 1/129),ns; **,ns
UAS-N-CA; UAS-Sprouty-RNAiBDSC 36709 + 520080% (n = 0/80) ,ns; **,ns

  1. Fisher’s Exact Test: ns = not significant.

  2. *

    Data from Table 1. ✝✝Another UAS-Egfr-RNAi line, BDSC 36773, did not show a phenotype with the control driver (C587-Gal4) and is not included here.

  3. †

    Compared with UAS-GFP RNAi.

  4. ‡

    Compared with UAS-EGFR Type II.

  5. §

    Compared with UAS-EGFR Type I; UAS-EGFR Type II.

  6. Compared with UAS-Sprouty RNAi.

  7. **

    Compared with UAS-EGFR Type I; UAS-Sprouty RNAi.

Table 3
Hub cell fate conversion after EGFR pathway activation.
Gal4 driverUAS lines (BDSC #)Days at 29 °C% Testes with GFP-marked cells outside the hub
E132-Gal4, Gal80tsUAS-G-TRACE (28280)80% (n = 0/39)
UAS-G-TRACE (28280); UAS-Egfr Type II (9533)61% (n = 19/31)****
UAS-G-TRACE (28280); UAS-Pointed.P1 (869)38% (n = 19/50)****
UAS-G-TRACE (28280); UAS-Pointed.P2 (399)22% (n = 4/18)**
UAS-G-TRACE (28280); UAS-Sprouty-RNAi (36709)36% (n = 32/89)****
UAS-G-TRACE (28280); UAS- Pten-RNAi (33643)18% (n = 6/34)**
E132-Gal4, Gal80tsUAS-G-TRACE (28281)80% (n = 0/84)
UAS-Egfr Type I (9534); UAS-G-TRACE (28281)2% (n = 1/47)ns

  1. Percentages in bold are significant compared to the negative control (corresponding UAS-G-TRACE alone).

  2. Fisher’s Exact Test: **p < 0.01, ****p < 0.0001, ns = not significant.

Table 4
Overexpression of EGF ligands in the adult testis niche does not cause hub cell proliferation.
Gal4 driverUAS lineSourceDays at 31 °C% Testes with PH3-positive hub cells
C587-Gal4, Gal80tsUAS-GFP-RNAiBDSC 9330 or 933170% (n = 0/294)*
UAS-secreted spitzBDSC 631347–90% (n = 0/219)a,ns
UAS-secreted spitzBDSC 5843670% (n = 0/75)a,ns
UAS-gurken ΔTCQueenan et al., 199970% (n = 0/82)a,ns
UAS-secreted gurkenBDSC 5841770% (n = 0/118)a,ns
UAS-secreted kerenUrban et al., 200270% (n = 0/113)a,ns
UAS-veinSchnepp et al., 199660% (n = 0/96)a,ns
E132-Gal4, Gal80tsUAS-GFP-RNAiBDSC 9330 or 933170% (n = 0/237)*
UAS-secreted spitzBDSC 6313470% (n = 0/208)a,ns
UAS-secreted spitzBDSC 5843670% (n = 0/92)a,ns
UAS-gurken ΔTCQueenan et al., 199970% (n = 0/86)a,ns
UAS-secreted gurkenBDSC 5841771% (n = 1/77)a,ns
UAS-secreted kerenUrban et al., 200270% (n = 0/113)a,ns
UAS-veinSchnepp et al., 1996------
C587-Gal4, Gal80tsUAS-secreted spitz +UAS-Argos-RNAiBDSC 63134 +BDSC 283837–92% (n = 4/203)b,ns

  1. Fisher’s Exact Test: ns = not significant (a, compared to UAS-GFP-RNAi control; b, compared to UAS-Argos-RNAi BDSC 28383 alone [Table 1]).

  2. *

    Data from Table 1.

Table 5
Ablation phenotypes with and without reduced EGFR.
GenotypeDays recoveredHub/GC/CySCHub/GCHub/CySCHub onlyNo hub
C587-Gal4; UAS-Grim/+; Tub-Gal80ts0< 1%(n = 2/213)93%(n = 197/213)< 1%(n = 1/213)4%(n = 8/213)2%(n = 5/213)
C587-Gal4; UAS-Grim/ EgfrF24; Tub-Gal80ts0*,§,ns3%(n = 3/108)87%(n = 94/108)0%(n = 0/108)8%(n = 9/108)2%(n = 2/108)
C587-Gal4; UAS-Grim/+; Tub-Gal80ts780%(n = 180/225)4%(n = 9/225)1%(n = 3/180)13%(n = 29/225)2%(n = 4/225)
C587-Gal4; UAS-Grim/ EgfrF24; Tub-Gal80ts7†,§,****45%(n = 64/142)20%(n = 28/142)< 1%(n = 1/142)34%(n = 48/142)< 1%(n = 1/142)
C587-Gal4; UAS-Grim/+; Tub-Gal80ts1481%(n = 123/151)1%(n = 2/151)2%(n = 3/151)15%(n = 22/151)< 1%(n = 1/151)
C587-Gal4; UAS-Grim/ EgfrF24; Tub-Gal80ts14‡,§,***55%(n = 64/116)6%(n = 7/116)3%(n = 3/116)33%(n = 39/116)3%(n = 3/116)
  1. *

    Compared with C587-Gal4; UAS-Grim/+; Tub Gal80ts at 0 days recovered.

  2. †

    Compared with C587-Gal4; UAS-Grim/+; Tub Gal80ts at 7 days recovered.

  3. ‡

    Compared with C587-Gal4; UAS-Grim/+; Tub Gal80ts at 14 days recovered.

  4. §

    Chi Square Test: ***p < 0.001, ****p < 0.0001, ns = not significant.

Table 6
Percentage of testes with ectopic hubs after 14 day recovery from CySC ablation.
GenotypeDaysRecovered% Recovered Testes with Ectopic Hubs% Recovered Testes without Ectopic Hubs
C587-Gal4; UAS-Grim/+; Tub-Gal80ts1429%(n = 139/487)71%(n = 348/487)
C587-Gal4; UAS-Grim/EgfrF24; Tub-Gal80ts14****12%(n = 32/270)88%(n = 238/270)

  1. Fisher’s Exact Test: ****p < 0.0001 (compared to testes with ectopic hubs in control C587-Gal4; UAS-Grim/+; Tub-Gal80ts flies).

Appendix 1—key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Genetic reagent (D. melanogaster)E132Gal4: P{w[ + mW.hs] = GawB}E132, w[*] (also called upd-Gal4)Bloomington Drosophila Stock CenterRRID:BDSC_26796
Genetic reagent (D. melanogaster)TubGal80ts: w[*];P{w[ + mC] = tubP-GAL80[ts]}2/TM2Bloomington Drosophila Stock CenterRRID:DSC_7017
Genetic reagent (D. melanogaster)UAS-GFP RNAi: w[1118]; P{w[ + mC] = UAS GFP.dsRNA.R}142Bloomington Drosophila Stock CenterRRID:BDSC_9330
Genetic reagent (D. melanogaster)UAS-GFP RNAi: w[1118]; P{w[ + mC] = UAS GFP.dsRNA.R}143Bloomington Drosophila Stock CenterRRID:BDSC_9331
Genetic reagent (D. melanogaster)UAS-Rbf RNAi: y1 sc[*] v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS03004}attP2/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_36744
Genetic reagent (D. melanogaster)UAS-Rbf RNAi: y1 sc[*] v1; P{y[ + t7.7] v[ + t1.8] = TRiP .GL01293}attP40/CyOBloomington Drosophila Stock CenterRRID:BDSC_41863
Genetic reagent (D. melanogaster)C587Gal4: P{w[ + mW.hs] = GawB}C587, w[*]Bloomington Drosophila Stock CenterRRID:BDSC_67747
Genetic reagent (D. melanogaster)UAS-Pointed.P1: w[1118]; P{w[ + mC] pnt[P1.UAS] = UAS pnt.P1}3Bloomington Drosophila Stock CenterRRID:BDSC_869
Genetic reagent (D. melanogaster)UAS-Pointed.P2: w[1118]; P{w[ + mC] pnt[P2.UAS] = UAS pnt.P2}2/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_399
Genetic reagent (D. melanogaster)UAS-Ras85D.V12: w[1118]; P{w[ + mC] = UAS-Ras85D.V12}TL1Bloomington Drosophila Stock CenterRRID:BDSC_4847
Genetic reagent (D. melanogaster)UAS-Rolled: y1 w[*]; P{w[ + mC] = UAS-rl[Sem].S}2Bloomington Drosophila Stock CenterRRID:BDSC_59006
Genetic reagent (D. melanogaster)UAS-Egfr Type I: w[*]; P{w[ + mC] = Egfr.1 .A887T.UAS}12–4/CyO, P{ry[ + t7.2] = sevRas1 .V12}FK1Bloomington Drosophila Stock CenterRRID:BDSC_9534
Genetic reagent (D. melanogaster)UAS-Egfr Type II: w[*]; P{w[ + mC] = Egfr.2 .A887T.UAS}8–2Bloomington Drosophila Stock CenterRRID:BDSC_9533
Genetic reagent (D. melanogaster)UAS-Egfrλ: w[*]; P{w[ + mC] = UAS Egfr.lambdatop}3/TM6C, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_59843
Genetic reagent (D. melanogaster)UAS-PVRλ: w[1118]; P{w[ + mC] = UASp Pvr.lambda}mP10Bloomington Drosophila Stock CenterRRID:BDSC_58496
Genetic reagent (D. melanogaster)UAS-PVRλ: w[1118]; P{w[ + mC] = UASp Pvr.lambda}mP1Bloomington Drosophila Stock CenterRRID:BDSC_58428
Genetic reagent (D. melanogaster)UAS-InR: y1 w[1118]; P{w[ + mC] = UAS InR.K414P}2Bloomington Drosophila Stock CenterRRID:BDSC_8250
Genetic reagent (D. melanogaster)UAS-InR: y1 w[1118]; P{w[ + mC] = UAS InR.A1325D}2Bloomington Drosophila Stock CenterRRID:BDSC_8263
Genetic reagent (D. melanogaster)UAS-Heartlessλ: y1 w[*]; P{w[ + mC] = UAS htl.lambda.M}40-22-2Bloomington Drosophila Stock CenterRRID:BDSC_5367
Genetic reagent (D. melanogaster)UAS-Breathlessλ: w[*]; P{w[ + mC] = UAS btl.lambda}2Bloomington Drosophila Stock CenterRRID:BDSC_29045
Genetic reagent (D. melanogaster)UAS-Sprouty RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01599}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36709
Genetic reagent (D. melanogaster)UAS-Pten RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00044}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33643
Genetic reagent (D. melanogaster)UAS-Argos RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.JF03020}attP2Bloomington Drosophila Stock CenterRRID:BDSC_28383
Genetic reagent (D. melanogaster)UAS-Egfr DN: y1 w[*]; P{w[ + mC] = UAS Egfr.DN.B}29-77-1; P{w[ + mC] = UAS Egfr.DN.B}29-8-1Bloomington Drosophila Stock CenterRRID:BDSC_5364
Genetic reagent (D. melanogaster)UAS-Egfr RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.JF02283}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36770
Genetic reagent (D. melanogaster)UAS-Egfr RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS05003}attP40Bloomington Drosophila Stock CenterRRID:BDSC_60012
Genetic reagent (D. melanogaster)UAS-Egfr RNAi: w1118; P{GD1654}v43267Vienna Drosophila Resource CenterStock #: 43,267
Genetic reagent (D. melanogaster)UAS-Egfr RNAi: P{KK100051}VIE-260BVienna Drosophila Resource CenterStock #: 107,130
Genetic reagent (D. melanogaster)UAS-N-CA: P{ry[ + t7.2] = hsFLP}1, y1 w[*]; P{w[ + mC] = UAS N.intra.GS}2/CyO; MKRS/TM2Bloomington Drosophila Stock CenterRRID:BDSC_52008No longer available
Genetic reagent (D. melanogaster)UAS-N-DN: y[ + t7.2] = hsFLP}12, y1 w[*]; P{w[ + mC] = UAS N.ECN}2; MKRS/TM2Bloomington Drosophila Stock CenterRRID:BDSC_51667
Genetic reagent (D. melanogaster)UAS-Notch RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00001}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33611
Genetic reagent (D. melanogaster)UAS-Notch RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00009}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33616
Genetic reagent (D. melanogaster)UAS-Notch RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00092}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35213
Genetic reagent (D. melanogaster)UAS-Notch RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.GLV21004}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35620
Genetic reagent (D. melanogaster)UAS-G-TRACE: w[*]; P{w[ + mC] = UAS-RedStinger}4, P{w[ + mC] = UAS FLP.D}JD1, P{w[ + mC] = Ubi-p63E(FRT.STOP)Stinger}9F6/CyOBloomington Drosophila Stock CenterRRID:BDSC_28280
Genetic reagent (D. melanogaster)UAS-G-TRACE: w[*]; P{w[ + mC] = UAS-RedStinger}6, P{w[ + mC] = UAS FLP.Exel}3, P{w[ + mC] = Ubi-p63E(FRT.STOP)Stinger}15F2Bloomington Drosophila Stock CenterRRID:BDSC_28281
Genetic reagent (D. melanogaster)UAS-secreted spitz: w[*]; P{w[ + mC] = UAS-sSpiCS}T28Bloomington Drosophila Stock CenterRRID:BDSC_63134
Genetic reagent (D. melanogaster)UAS-secreted spitz: w[*]; P{w[ + mC] = UASp spi.sec}3/TM3, Ser1()Bloomington Drosophila Stock CenterRRID:BDSC_58436
Genetic reagent (D. melanogaster)UAS-secreted gurken: w[*]; P{w[ + mC] = UASp grk.sec}2/CyOBloomington Drosophila Stock CenterRRID:BDSC_58417
Genetic reagent (D. melanogaster)UAS-gurken ΔTCPMID: 10559478Dr. Trudi Schüpbach (Princeton University)
Genetic reagent (D. melanogaster)UAS- secreted KerenPMID:12169630Dr. Matthew Freeman (MRC Laboratory of Molecular Biology)
Genetic reagent (D. melanogaster)UAS-VeinPMID:8824589Dr. Amanda Simcox (The Ohio State University)
Genetic reagent (D. melanogaster)UAS-GrimPMID:9846179Dr. John Nambu (University of Massachusetts)
Genetic reagent (D. melanogaster)Egfr(-): Egfr[f24]/T(2;3)TSTL, CyO: TM6B, Tb1Bloomington Drosophila Stock CenterRRID:BDSC_6500
Antibody(Mouse monoclonal) anti–Fasciclin III (Drosophila)DSHBCat#: 7G10; RRID: AB_528238IHC (1:50)
Antibody(Mouse monoclonal) anti-phospho-Histone H3 (Ser10) (6G3)Cell Signaling TechnologyCat#: 9,706 S; RRID:AB_331748IHC (1:400)
Antibody(Guinea Pig polyclonal) anti-Traffic JamLaboratory of D. Godt (Li et al., 2003)N/AIHC (1:20,000)
Antibody(Rabbit polyclonal) anti-Vasa (d-260)Santa Cruz BiotechnologyCat#: SC-30210; RRID:AB_793874IHC (1:200)
Antibody(Chicken polyclonal) anti-GFPAbcamCat#: ab13970; RRID:AB_300798IHC (1:10,000)
Antibody(Rabbit polyclonal) anti-DsRedTakara BioCat#: 632496; RRID:AB_10013483IHC (1:10,000)
Antibody(Rabbit polyclonal) anti-dpErkCell Signaling TechnologyCat#: 4370; RRID:AB_2315112IHC (1:100)
Antibody(Goat polyclonal) anti-Mouse IgG (H + L) secondary antibody, Alexa Fluor 488 conjugateThermoFisher ScientificCat#: A11029; RRID:AB_2534088IHC (1:400)
Antibody(Goat polyclonal) anti-Rabbit IgG (H + L) secondary antibody, Alexa Fluor 568 conjugateThermoFisher ScientificCat#: A11011; RRID:AB_143157IHC (1:200)
Antibody(Goat polyclonal) anti-Chicken IgY (H + L) secondary antibody, Alexa Fluor 488 conjugateThermoFisher ScientificCat#: A11039; RRID:AB_2534096IHC (1:400)
Antibody(Goat polyclonal) anti-Guinea Pig IgG (H + L) secondary antibody, Alexa Fluor 568 conjugateThermoFisher ScientificCat#: A11075; RRID:AB_2534119IHC (1:200)
Antibody(Goat polyclonal) anti-Guinea Pig IgG (H + L) secondary antibody, Alexa Fluor 633 conjugateThermoFisher ScientificCat#: A21105; RRID:AB_2535757IHC (1:200)
Chemical compound, drug4,6-diamidino-2-phenylindole (DAPI)Millipore/Sigma (formerly Sigma-Aldrich)Cat#: 10236276001; CAS: 28718-90-3IHC (1 μg/mL)
Chemical compound, drug16% Paraformaldehyde (formaldehyde) aqueous solutionElectron Microscopy SciencesCat#: 15710; CAS: 50-00-0
Chemical compound, drugGoat SerumMillipore/Sigma (formerly Sigma-Aldrich)Cat#: G9023
Chemical compound, drugVectashield antifade mounting mediumVector LaboratoriesCat#: H-1000
Chemical compound, drugPhosphatase Inhibitor Cocktail 2Millipore/Sigma (formerly Sigma-Aldrich)Cat#: P5726
Chemical compound, drugDMSO (Dimethyl Sulfoxide), SterileCell Signaling TechnologyCat#: 12,611 P
Software, algorithmFijiSchindelin et al., 2012https://www.fiji.sc/
Software, algorithmZeiss LSMCarl Zeiss Microscopyhttps://www.zeiss.com/microscopy/us/downloads/lsm-5-series.html
Software, algorithmZenCarl Zeiss Microscopyhttps://www.zeiss.com/microscopy/int/products/microscope-software/zen.html
Software, algorithmPrism 6GraphPadhttp://www.graphpad.com/scientific-software/prism/
Appendix 1—table 1
Screen Summary.
Reagent type (species) or resourceDesignationSource or referenceIdentifiersPathwayAdditionalInformation
Genetic reagent (D. melanogaster)UAS-EcR-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC03114}attP2/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_50712Ecdysone
Genetic reagent (D. melanogaster)UAS-Ecr RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMJ22371}attP40Bloomington Drosophila Stock CenterRRID:BDSC_58286Ecdysone
Genetic reagent (D. melanogaster)UAS-btl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS02038}attP2Bloomington Drosophila Stock CenterRRID:BDSC_ 40871FGFR
Genetic reagent (D. melanogaster)UAS-btl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS02656}attP40Bloomington Drosophila Stock CenterRRID: BDSC_43544FGFR
Genetic reagent (D. melanogaster)UAS-btl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC04140}attP2Bloomington Drosophila Stock CenterRRID:BDSC_55870FGFR
Genetic reagent (D. melanogaster)UAS-btl-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS05005}attP40Bloomington Drosophila Stock CenterRRID:BDSC_ 60013FGFR
Genetic reagent (D. melanogaster)UAS-htl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01437}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35024FGFR
Genetic reagent (D. melanogaster)UAS-htl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS04514}attP40Bloomington Drosophila Stock CenterRRID:BDSC_57313FGFR
Genetic reagent (D. melanogaster)UAS-htl-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMJ22375}attP40Bloomington Drosophila Stock CenterRRID:BDSC_ 58289FGFR
Genetic reagent (D. melanogaster)UAS-Galphaf-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL01545}attP40Bloomington Drosophila Stock CenterRRID:BDSC_43201GPCR
Genetic reagent (D. melanogaster)UAS-Galphai-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01273}attP2/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_34924GPCR
Genetic reagent (D. melanogaster)UAS-Galphai-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00328}attP2Bloomington Drosophila Stock CenterRRID:BDSC_ 35407GPCR
Genetic reagent (D. melanogaster)UAS-Galphai-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS02138}attP40Bloomington Drosophila Stock CenterRRID:BDSC_40890GPCR
Genetic reagent (D. melanogaster)UAS-Galphao-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01129}attP2Bloomington Drosophila Stock CenterRRID:BDSC_34653GPCR
Genetic reagent (D. melanogaster)UAS-Galphaq-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.JF02464}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33765GPCR
Genetic reagent (D. melanogaster)UAS-Galphaq-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS03015}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36775GPCR
Genetic reagent (D. melanogaster)UAS-Galphaq-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL01048}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36820GPCR
Genetic reagent (D. melanogaster)UAS-Galphas-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC03106}attP2Bloomington Drosophila Stock CenterRRID:BDSC_50704GPCR
Genetic reagent (D. melanogaster)UAS-ci: w[*]; P{w[ + mC] = UAS ci.HA.wt}3Bloomington Drosophila Stock CenterRRID:BDSC_32570Hedgehog
Genetic reagent (D. melanogaster)UAS-Ci-activated: w[*]; P{w[ + mC] = UAS HA.ci.m1-3*103}2Bloomington Drosophila Stock CenterRRID:BDSC_32571Hedgehog
Genetic reagent (D. melanogaster)UAS-ci-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC05801}attP2Bloomington Drosophila Stock CenterRRID:BDSC_64928Hedgehog
Genetic reagent (D. melanogaster)UAS-Hmgcr-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC03053}attP40Bloomington Drosophila Stock CenterRRID:BDSC_50652Hedgehog
Genetic reagent (D. melanogaster)UAS-hpo-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00006}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33614Hippo
Genetic reagent (D. melanogaster)UAS-hpo-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00046}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35176Hippo
Genetic reagent (D. melanogaster)UAS-yki-activated: w[*]; P{y[ + t7.7] w[ + mC] = UAS yki.S111A.S168A.S250A.V5}attP2Bloomington Drosophila Stock CenterRRID:BDSC_28817Hippo
Genetic reagent (D. melanogaster)UAS-yki-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00041}attP2Bloomington Drosophila Stock CenterRRID:BDSC_34067Hippo
Genetic reagent (D. melanogaster)UAS-Akt: y1 w[1118]; P{w[ + mC] = UAS Akt.Exel}2Bloomington Drosophila Stock CenterRRID:BDSC_8191InR
Genetic reagent (D. melanogaster)UAS-Akt-activated: w[*]; P{w[ + mC] = UAS-myr-Akt1.V}3/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_50758InRNo longer available
Genetic reagent (D. melanogaster)UAS-Akt-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00007}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33615InR
Genetic reagent (D. melanogaster)UAS-Akt-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.JF02668}attP2Bloomington Drosophila Stock CenterRRID:BDSC_27518InR
Genetic reagent (D. melanogaster)UAS-Foxo: y1 w[*]†; P{w[ + mC] = UAS foxo.P}2Bloomington Drosophila Stock CenterRRID:BDSC_9575InR
Genetic reagent (D. melanogaster)UAS-Foxo: w[1118]; P{w[ + mC] = UASp foxo.S}3Bloomington Drosophila Stock CenterRRID:BDSC_42221InR
Genetic reagent (D. melanogaster)UAS-Foxo: w[1118]; P{w[ + mC] = UASp foxo.GFP}3Bloomington Drosophila Stock CenterRRID:BDSC_43633InR
Genetic reagent (D. melanogaster)UAS-Foxo: w[1118]; P{w[ + mC] = UASp foxo.GFP}2Bloomington Drosophila Stock CenterRRID:BDSC_44214InR
Genetic reagent (D. melanogaster)UAS-Foxo RNAi: y[1] sc[*] v[1] sev[21]; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00422}attP2Bloomington Drosophila Stock CenterRRID:BDSC_32427InR
Genetic reagent (D. melanogaster)UAS-Foxo RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00793}attP2Bloomington Drosophila Stock CenterRRID:BDSC_32993InR
Genetic reagent (D. melanogaster)UAS-InR-DN: y1 w[1118]; P{w[ + mC] = UAS InR.K1409A}2Bloomington Drosophila Stock CenterRRID:BDSC_8252InR
Genetic reagent (D. melanogaster)UAS-InR-DN: y1 w[1118]; P{w[ + mC] = UAS InR.K1409A}3Bloomington Drosophila Stock CenterRRID:BDSC_8253InR
Genetic reagent (D. melanogaster)UAS-InR-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS03166}attP40Bloomington Drosophila Stock CenterRRID:BDSC_51518InR
Genetic reagent (D. melanogaster)UAS-Pi3K21B: y1 w[*]; P{w[ + mC] = UAS-Pi3K21B.HA}2Bloomington Drosophila Stock CenterRRID:BDSC_25899InR
Genetic reagent (D. melanogaster)UAS-dome-RNAi: w[1118]; P{GD14494}v36356Vienna Drosophila Resource CenterStock #: 36356Jak-Stat
Genetic reagent (D. melanogaster)UAS-dome-RNAi:P{KK104700}VIE-260BVienna Drosophila Resource CenterStock #: 106071Jak-Stat
Genetic reagent (D. melanogaster)UAS-hop-activated: w; UAS-hop[TumL]/CyOPMID:7796812Jak-StatDr. Norbert Perrimon(Harvard Medical School)
Genetic reagent (D. melanogaster)UAS-Stat92E-RNAi: w[1118]; P{GD4492}v43866Vienna Drosophila Resource CenterStock #: 43866Jak-Stat
Genetic reagent (D. melanogaster)UAS-Stat92E-RNAi:P{KK100519}VIE-260BVienna Drosophila Resource CenterStock #: 106980Jak-Stat
Genetic reagent (D. melanogaster)UAS-upd1PMID: 10346822Jak-StatDr. David Strutt (Harvard Medical School)
Genetic reagent (D. melanogaster)UAS-upd1-RNAi: w[1118]; P{GD1158}v3282Stock #: 3282Jak-Stat
Genetic reagent (D. melanogaster)UAS-upd1-RNAi: y[1] sc[*] v[1] sev[21]; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00545}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33680Jak-Stat
Genetic reagent (D. melanogaster)UAS-upd2-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00901}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33949Jak-Stat
Genetic reagent (D. melanogaster)UAS-upd2-RNAi:1sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00948}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33988Jak-Stat
Genetic reagent (D. melanogaster)UAS-upd3-RNAi: y1 sc[ *]‡ v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00646}attP2Bloomington Drosophila Stock CenterRRID:BDSC_32859Jak-Stat
Genetic reagent (D. melanogaster)UAS-hep-activated: y1 w[1118]; P{w[ + mC] = UAS hep.CA}4Bloomington Drosophila Stock CenterRRID:BDSC_6406Jnk
Genetic reagent (D. melanogaster)UAS-hep-activated: w[*]; P{w[ + mC] = UAS Hep.Act}2Bloomington Drosophila Stock CenterRRID:BDSC_9306Jnk
Genetic reagent (D. melanogaster)UAS-kay: w[1118]; P{w[ + mC] = UAS-Fra}2Bloomington Drosophila Stock CenterRRID:BDSC_7213Jnk
Genetic reagent (D. melanogaster)UAS-kay-DN: w[1118]; P{w[ + mC] = UAS Fra.Fbz}5Bloomington Drosophila Stock CenterRRID:BDSC_7214Jnk
Genetic reagent (D. melanogaster)UAS-kay-DN: y1 w[1118]; P{w[ + mC] = UAS Fra.Fbz}7Bloomington Drosophila Stock CenterRRID:BDSC_7215Jnk
Genetic reagent (D. melanogaster)UAS-jra: y1 w[1118]; P{w[ + mC] = UAS-Jra}2Bloomington Drosophila Stock CenterRRID:BDSC_7216Jnk
Genetic reagent (D. melanogaster)UAS-kay-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00254}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33379Jnk
Genetic reagent (D. melanogaster)UAS-wgn-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC03962}attP40Bloomington Drosophila Stock CenterRRID:BDSC_55275Jnk
Genetic reagent (D. melanogaster)UAS-egr-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC03963}attP40Bloomington Drosophila Stock CenterRRID:BDSC_55276Jnk
Genetic reagent (D. melanogaster)UAS-Pvr-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01662}attP40Bloomington Drosophila Stock CenterRRID:BDSC_37520Pvr
Genetic reagent (D. melanogaster)UAS-Pvf1-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01958}attP40Bloomington Drosophila Stock CenterRRID:BDSC_39038Pvr
Genetic reagent (D. melanogaster)UAS-Pvr-DN: w[1118]; P{w[ + mC] = UASp Pvr.DN}D1/CyOBloomington Drosophila Stock CenterRRID:BDSC_58430Pvr
Genetic reagent (D. melanogaster)UAS-Pvr-DN: w[1118]; P{w[ + mC] = UASp Pvr.DN}D7Bloomington Drosophila Stock CenterRRID:BDSC_58431Pvr
Genetic reagent (D. melanogaster)UAS-aop: w[*]; P{w[ + mC] = UAS aop.WT}Ia/CyOBloomington Drosophila Stock CenterRRID:BDSC_5790RTK
Genetic reagent (D. melanogaster)UAS-aop-activated: w[*]; P{w[ + mC] = UAS aop.ACT}IIaBloomington Drosophila Stock CenterRRID:BDSC_5789RTK
Genetic reagent (D. melanogaster)UAS-pnt-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01452}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35038RTK
Genetic reagent (D. melanogaster)UAS-rl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00173}attP2Bloomington Drosophila Stock CenterRRID:BDSC_34855RTK
Genetic reagent (D. melanogaster)UAS-rl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00215}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36058RTK
Genetic reagent (D. melanogaster)UAS-sev-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.JF02393}attP2Bloomington Drosophila Stock CenterRRID:BDSC_36778RTK
Genetic reagent (D. melanogaster)UAS-sev-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMC04136}attP2Bloomington Drosophila Stock CenterRRID:BDSC_55866RTK
Genetic reagent (D. melanogaster)UAS-tor-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00021}attP2Bloomington Drosophila Stock CenterRRID:BDSC_33627RTK
Genetic reagent (D. melanogaster)UAS-tor-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP.HMJ22419}attP40Bloomington Drosophila Stock CenterRRID:BDSC_58312RTK
Genetic reagent (D. melanogaster)UAS-Mad:P{ry[ + t7.2] = hsFLP}12, y1 w[*]; P{w[ + mC] = UAS Mad.FLAG}2; P{y[ + t7.7] w[ + mC] = mir-ban-lacZ.brC12}attP2Bloomington Drosophila Stock CenterRRID:BDSC_44256TGFβ
Genetic reagent (D. melanogaster)UAS-Mad-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.GLV21013}attP2/TM3, Sb1Bloomington Drosophila Stock CenterRRID:BDSC_35648TGFβ
Genetic reagent (D. melanogaster)UAS-Mad-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL01527}attP40Bloomington Drosophila Stock CenterRRID:BDSC_43183TGFβ
Genetic reagent (D. melanogaster)UAS-Smox-RNAi:1sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS02203}attP40Bloomington Drosophila Stock CenterRRID:BDSC_41670TGFβ
Genetic reagent (D. melanogaster)UAS-Smox-RNAi: y1 v1; P{y[ + t7.7] v[ + t1.8] = TRiP .GL01476}attP2Bloomington Drosophila Stock CenterRRID:BDSC_43138TGFβ
Genetic reagent (D. melanogaster)UAS-cact-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00084}attP2Bloomington Drosophila Stock CenterRRID:BDSC_34775Toll
Genetic reagent (D. melanogaster)UAS-cact-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00627}attP40Bloomington Drosophila Stock CenterRRID:BDSC_37484Toll
Genetic reagent (D. melanogaster)UAS-dl: y1 w[*]; P{w[ + mC] = UAS dl.H}2Bloomington Drosophila Stock CenterRRID:BDSC_9319Toll
Genetic reagent (D. melanogaster)UAS-dl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00727}attP2Bloomington Drosophila Stock CenterRRID:BDSC_32934Toll
Genetic reagent (D. melanogaster)UAS-dl-RNAi:y1sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS00028}attP2Bloomington Drosophila Stock CenterRRID:BDSC_34938Toll
Genetic reagent (D. melanogaster)UAS-dl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00610}attP40Bloomington Drosophila Stock CenterRRID:BDSC_36650Toll
Genetic reagent (D. melanogaster)UAS-dl-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP .GL00676}attP2Bloomington Drosophila Stock CenterRRID:BDSC_38905Toll
Genetic reagent (D. melanogaster)UAS-arm-RNAi: y1 sc[*] v1 sev21; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS01414}attP2Bloomington Drosophila Stock CenterRRID:BDSC_35004Wnt
Genetic reagent (D. melanogaster)UAS-pan: y1 w[1118]; P{w[ + mC] = UAS pan.dTCF}24/CyOBloomington Drosophila Stock CenterRRID:BDSC_4837Wnt
Genetic reagent (D. melanogaster)UAS-pan: y1 w[1118]; P{w[ + mC] = UAS pan.dTCF}4Bloomington Drosophila Stock CenterRRID:BDSC_4838Wnt
Genetic reagent (D. melanogaster)UAS-pan-constitutive repressor: y[1] w[1118]; P{w[ + mC] = UAS pan.dTCFDeltaN}4Bloomington Drosophila Stock CenterRRID:BDSC_4784Wnt
Genetic reagent (D. melanogaster)UAS-pan-constitutive repressor: y[1] w[1118]; P{w[ + mC] = UAS pan.dTCFDeltaN}5Bloomington Drosophila Stock CenterRRID:BDSC_4785Wnt
Genetic reagent (D. melanogaster)UAS-pan-RNAi: y v; P{y[ + t7.7] v[ + t1.8] = TRiP.HMS02015}attP40/CyOBloomington Drosophila Stock CenterRRID:BDSC_40848Wnt
Genetic reagent (D. melanogaster)UAS-wg-RNAi: w[1118]; P{GD5007}v13352Vienna Drosophila Resource CenterStock #: 13352Wnt
  1. *

    n = at least 20–199 testes for all lines.

  2. †

    lines listed in the key resource table are not repeated here.

  3. ‡

    no lines listed here had significant numbers of dividing hub cells compared to UAS-GFP-RNAi controls.

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  1. Leah J Greenspan
  2. Margaret de Cuevas
  3. Kathy H Le
  4. Jennifer M Viveiros
  5. Erika L Matunis
(2022)
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