1. Cell Biology
  2. Developmental Biology

Protein phosphatase 1 activity controls a balance between collective and single cell modes of migration

  1. Yujun Chen
  2. Nirupama Kotian
  3. George Aranjuez
  4. Lin Chen
  5. C Luke Messer
  6. Ashley Burtscher
  7. Ketki Sawant
  8. Damien Ramel
  9. Xiaobo Wang
  10. Jocelyn A McDonald  Is a corresponding author
  1. Division of Biology, Kansas State University, United States
  2. Lerner Research Institute, Cleveland Clinic, United States
  3. LBCMCP, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, France
https://doi.org/10.7554/eLife.52979
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Cite this article
  1. Yujun Chen
  2. Nirupama Kotian
  3. George Aranjuez
  4. Lin Chen
  5. C Luke Messer
  6. Ashley Burtscher
  7. Ketki Sawant
  8. Damien Ramel
  9. Xiaobo Wang
  10. Jocelyn A McDonald
(2020)
Protein phosphatase 1 activity controls a balance between collective and single cell modes of migration
eLife 9:e52979.
https://doi.org/10.7554/eLife.52979
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8 figures, 17 videos, 3 tables and 1 additional file

Figures

Figure 1 with 3 supplements
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NiPp1 expression causes the border cell cluster to fall apart and disrupts migration.

(A–F) Wild-type border cell migration during oogenesis stages 9 and 10. (A–C) Egg chambers at the indicated stages labeled with E-Cadherin (E-Cad; green), F-actin (magenta) and DAPI (blue). …

Figure 1—figure supplement 1
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Patterns of GAL4s expressed in border cells.

Expression patterns of c306-GAL4 (A) and slbo-GAL4 (B) during oogenesis, indicated by UAS-GFP.nls (green; arrowheads, brackets). All egg chambers were co-stained with DAPI (blue) to label nuclei and …

Figure 1—figure supplement 2
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Cell-specific phenotypes induced by NiPp1.

(A) Fertility of control versus NiPp1-expressing females. The average progeny per female in each vial (individual plot points) is shown as a box-and-whiskers plot (see Figure 1 legend for details of …

Figure 1—figure supplement 3
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NiPp1 does not greatly alter border cell specification or cell number per cluster.

(A–B’) Stage 10 wild-type (A, A’) and NiPp1-expressing (B, B’) egg chambers stained with anti-Slbo (green in A, B; white in A’, B’), anti-Eya (red in A, B), and DAPI to detect nuclei (blue in A, B). …

Figure 2 with 2 supplements
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Pp1c expression in border cells and specificity of NiPp1 inhibition of Pp1c activity.

(A–F) Stage 9 and 10 egg chambers showing the endogenous patterns of Pp1c subunits (green) in border cells (arrowheads), follicle cells, and the germline nurse cells and oocyte. DAPI (blue) labels …

Figure 2—figure supplement 1
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Rescue of NiPp1 phenotypes by Pp1c genes.

(A–E) Overexpression of Pp1c subunits on their own does not impair border cell migration to the oocyte. Stage 10 egg chambers of the indicated genotypes stained for Armadillo (Arm; β-Catenin) to …

Figure 2—figure supplement 2
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NiPp1 promotes nuclear localization of Pp1c subunits.

(A–A”) NiPp1-HA overexpression alone localizes mainly in the nuclei, revealed by HA antibody (red in A, A’) and human NiPp1 antibody (PPP1R8; green in A, A’’). (B–C”) NiPp1-HA overexpression …

Figure 3 with 1 supplement
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Pp1c genes are required for normal border cell migration and cluster cohesion.

(A–F) Knockdown of Pp1c genes by RNAi disrupts border cell cluster migration and cohesion. (A–D) Stage 10 egg chambers expressing RNAi against the indicated genes were stained for SN (red) to label …

Figure 3—figure supplement 1
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Delamination and migration defects caused by loss of Pp1c.

(A) Quantification of live split border cell cluster parts that completed migration during the movie, for matched control and RNAi knockdown of the indicated Pp1c genes. N = 14 videos for control, n …

Figure 4 with 1 supplement
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The cadherin-catenin complex is required for the collective cohesion of the migrating border cell cluster and is regulated by Pp1.

(A–J) Knocking down E-Cad, β-Cat or α-Cat by RNAi disrupts border cell cluster migration and cohesion. Images of stage 10 egg chambers stained for phalloidin to label F-actin (red) and DAPI to label …

Figure 4—figure supplement 1
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RNAi for cadherin-catenin reduces endogenous levels of the specifically targeted gene.

(A–H’) Efficiency of cadherin-catenin RNAi in border cells as detected by antibody staining to the respective proteins. Stage 10 control (A, A’, C, C’, E, E’), E-Cad-RNAi (B, B’), β-Cat-RNAi (line …

Figure 5 with 1 supplement
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Pp1c is required for normal border cell protrusion dynamics.

(A–B””) Frames from a matched control (Video 6; A–A””) and a Pp1α-96A-RNAi (Video 8; B–B””) showing the migrating border cell cluster expressing the membrane marker PLCδ-PH-EGFP. Time in min. Arrows …

Figure 5—figure supplement 1
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Additional quantification of protrusion dynamics and Rac activity in Pp1-inhibited and α-Cat-RNAi border cells.

(A) Close-up view of a live border cell cluster depicting how protrusions are measured. The main body of the border cell cluster is outlined (yellow circle). The protrusion length and area (green …

Figure 6 with 3 supplements
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Pp1 activity promotes normal border cell shape and distribution of actomyosin in the border cell cluster.

(A–C) Pp1 is required for border cell shape. (A–B’) Examples of control (A, A’) and NiPp1-expressing border cells (B, B’). Cell shape was visualized using the membrane marker PLCδ-PH-EGFP driven by s…

Figure 6—figure supplement 1
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Pp1 restricts the distribution of Myo-II to the cluster periphery during early migration.

(A–B’’’’’) Stills from representative confocal videos of dynamic Sqh-GFP in early-migration borders cells over the course of 20 min. Image gain and other acquisition parameters were the same, except …

Figure 6—figure supplement 2
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RNAi for cadherin-catenin alters the actomyosin pattern of the border cell cluster.

(A, A’) Control mCherry-RNAi border cell cluster has higher F-actin (magenta arrows) at the periphery of the cluster, but lower levels inside (yellow arrow) the group. (B–D’) Knocking down E-Cad (B, …

Figure 6—figure supplement 3
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Myo-II is not required for cadherin-catenin enrichment at border cell-border cell contacts.

(A–D) Representative images showing the E-Cad (white in A, B; green in A’, B”) and β-cat (white in C, D; green in C’, D”) protein expression patterns in control (A–A’’, C–C’’) and sqh-RNAi (B–B’’, …

Figure 7 with 2 supplements
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Pp1, through myosin phosphatase, promotes contractility of the cluster.

(A–B’’) Pp1 restricts Myo-II activation to the cluster periphery. Representative images showing p-Sqh localization (white in A, B; red in A’’, (B’’) and membrane GFP (PLCδ-PH-EGFP; green in A’, (A’’,…

Figure 7—figure supplement 1
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Expression of Mbs during border cell migration and specificity of Mbs-RNAi knockdown.

(A–J) Mbs transcript (A–C) and protein (D–I”) are found in border cells throughout migration (arrowheads, A–F). (A–C) Mbs mRNA pattern as detected by in situ hybridization (green) in stage 9 to 10 …

Figure 7—figure supplement 2
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Pp1 promotes moderate levels of RhoA activity in border cells.

(A–B’) Representative processed Rho-FRET images in control (A, A’) and NiPp1 overexpressing (B, B’) border cells. The CFP channel (A, B) is shown. The FRET images (A’, B’) are color-coded as …

Figure 8
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Model for the Pp1 function in border cell migration.

(A) Schematic of the phenotypes and the localizations of Pp1c, F-actin, p-Sqh, and the cadherin-catenin complex during normal and Pp1-inhibited (NiPp1 expression or Pp1c-RNAi) border cell cluster …

Videos

Video 1
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Control (c306-GAL4/+; UAS-mCherry-Jupiter/+) egg chamber showing normal border cell migration.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 2
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NiPp1 overexpressing (c306-GAL4/+; UAS-mCherry-Jupiter/+; UAS-NiPp1/+) egg chamber showing the migration defect and splitting phenotype.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 3
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Representative time-lapse video of a stage 9 NiPp1 overexpressing (c306-GAL4,tsGAL80/+; UAS-mCherry-Jupiter/+; UAS-NiPp1/+) egg chamber with DIC channel.

Frames were acquired every 2 min with a 20x objective. Time is in hr:min. Anterior is to the left.

Video 4
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Representative time-lapse video of a stage 9 NiPp1 overexpressing (c306-GAL4,tsGAL80/+; UAS- mCherry-Jupiter /+;UAS-NiPp1/+) egg chamber with DIC channel.

Frames were acquired every 2 min with a 20x objective. Time is in hr:min. Anterior is to the left.

Video 5
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NiPp1 overexpressing (slbo-GAL4/+; UAS- PLCδ-PH-EGFP/UAS-NiPp1) egg chamber showing the loss of a membrane attachment between one border cell and the rest of the border cell cluster.

Anterior is to the left.

Video 6
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Control (c306-GAL4,tsGAL80/+; UAS- PLCδ-PH-EGFP/+) egg chamber showing normal border cell migration.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 7
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Representative time-lapse video of a stage 9 Pp1α−96A RNAi (c306-GAL4,tsGAL80/+; v27673/+; PLCδ-PH-EGFP /+) egg chamber.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 8
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Another representative time-lapse video of a stage 9 Pp1α−96A RNAi (c306-GAL4,tsGAL80/+; v27673/+;UAS-PLCδ-PH-EGFP/+) egg chamber.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 9
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Representative time-lapse video of a stage 9 Pp1-13C RNAi (c306-GAL4,tsGAL80/+;v29058/+;UAS-PLCδ-PH-EGFP/+) egg chamber.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 10
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Representative time-lapse video of a stage 9 Pp1-87B RNAi (c306-GAL4,tsGAL80/+; v35024/+;UAS-PLCδ-PH-EGFP/+) egg chamber.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 11
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Representative time-lapse video of a stage 9 α-Cat RNAi (c306-GAL4,tsGAL80/+; v107298/+; UAS-PLCδ-PH-EGFP/+) egg chamber.

Frames were acquired every 3 min with a 20x objective. Anterior is to the left.

Video 12
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Control (LifeAct-GFP/+) egg chamber showing the dynamics of F-actin with LifeAct-GFP, Frames were acquired every 2 min with a 40x water immersion objective.

We observed similar dynamics in three videos. Anterior is to the left.

Video 13
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NiPp1 overexpressing (slbo-Gal4/+; UAS-NiPp1/LifeAct-GFP) egg chamber showing F-actin dynamics with LifeAct-GFP.

Frames were acquired every 2 min with a 40x water immersion objective. We observed similar dynamics in three videos. Anterior is to the left.

Video 14
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Control (c306-GAL4, tsGAL80/+; Sqh-GFP/+) egg chamber showing normal Sqh-GFP dynamics in early migration.

Frames were acquired every 1 min with a 40x water immersion objective, only a single focal plane is shown, with a 3D projection of the entire z-stack at the beginning and the end of the video. …

Video 15
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Representative NiPp1 overexpressing (c306-GAL4, tsGAL80/+; UAS-NiPp1/Sqh-GFP) egg Chamber showing the Sqh-GFP dynamics in early migration.

Frames were acquired every 1 min with a 40x water immersion objective, only a single focal plane is shown, with a 3D projection of the entire z-stack at the beginning and the end of the video. …

Video 16
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Control (Sqh-GFP/+) egg chamber showing normal Sqh-GFP dynamics in mid-migration.

Frames were acquired every 1 min with a 40x water immersion objective, only a single focal plane is shown, with a 3D projection of the entire z-stack at the beginning and the end of the video. …

Video 17
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Representative NiPp1 overexpressing (c306-GAL4, tsGAL80/+; UAS-NiPp1/Sqh-GFP) egg chamber showing the Sqh-GFP dynamics in mid-migration.

Frames were acquired every 1 min with a 40x water immersion objective, only a single focal plane is shown, with a 3D projection of the entire z-stack at the beginning and the end of the video. …

Tables

Table 1
Summary of the PPP family screen.

Results of the targeted serine-threonine protein phosphatase RNAi screen.

Gene symbolAnnotation symbolRNAi lineMigration defect (c306-Gal4)Expression level in
ovary (modENCODE)
Pp2B-14DCG9842BDSC:25929Nomoderate
BDSC:40872No
VDRC:46873No
mtsCG7109BDSC:27723Pupal lethalmoderate
BDSC:38337No
BDSC:57034No
BDSC:60342No
Pp4-19CCG32505BDSC:27726Pupal lethalmoderate
BDSC:38372No
BDSC:57823Pupal lethal
VDRC:25317Yes
CanA-14FCG9819BDSC:38966Nomoderate
PpD3CG8402BDSC:57307Nomoderate
PpVCG12217BDSC:57765Nomoderate
NiPp1CG8980BDSC:23711Yesmoderate
CanA1CG1455BDSC:25850Nolow
CG11597CG11597BDSC:57047Novery low
BDSC:61988No
rgdCCG44746BDSC:60076Novery low
Table 2
Genotypes for figures.

List of genotypes shown in the figures.

FigurePanelGenotype
Figure 1A-Fw1118
Gc306-GAL4,tsGAL80/+
Hc306-GAL4,tsGAL80/+;UAS-NiPp1/+
Kc306-GAL4/+; UAS-Cherry:Jupiter / +
Lc306-GAL4/+; UAS-Cherry:Jupiter / +;UAS-NiPp1/+
Figure 1—figure supplement 1Ac306-GAL4,tsGAL80/+;UAS-GFP.nls/+
Bslbo-GAL4/+;UAS-GFP.nls/+
Figure 1—figure supplement 2Bc306-GAL4/+;UAS-PLCdelta-PH-EGFP/+
D-Gslbo-GAL4,UAS-mCD8-GFP/+;
slbo-GAL4,UAS-mCD8-GFP/+;UAS-NiPp1/+
H-Kupd-GAL4/+;UAS-mCD8.ChRFP/+
upd-GAL4/+;UAS-NiPp1/+
L-Nc306-GAL4,tsGAL80/+
c306-GAL4,tsGAL80/+;UAS-NiPp1/+
Figure 1—figure supplement 3Ac306-GAL4/+ (WT)
Bc306-GAL4,tsGAL80/+;UAS-NiPp1/+
Figure 2A-CFlyFos021765(pRedFlp-Hgr) (Pp1alpha-96A15346::2XTY1-SGFP-V5-preTEV-BLRP-3XFLAG)dFRT
D-Fw[1118] PBac{681 .P.FSVS-1}flw[CPTI002264]
G-Hc306-GAL4,tsGAL80/+;UAS-NiPp1/UAS-mCD8.ChRFP
c306-GAL4,tsGAL80/+;UAS-NiPp1/UAS-Pp1α−96A.HA
c306-GAL4,tsGAL80/+;UAS-NiPp1/UAS-Pp1-87B.HA
c306-GAL4,tsGAL80/+;UAS-NiPp1/UAS-Pp1-13C.HA
c306-GAL4,tsGAL80/+;UAS-NiPp1/UAS-Flw.3xHA
c306-GAL4,tsGAL80/+;UAS-hPPP1CC/+;UAS-NiPp1/
Figure 2—figure supplement 1Ac306-GAL4/+;UAS-Pp1α−96A.HA/+
Bc306-GAL4/+;UAS-Pp1-87B.HA/+
Cc306-GAL4/+;UAS-Pp1-13C.HA/+
Dc306-GAL4/+;UAS-Flw.3xHA/+
Ec306-GAL4/+;UAS-hPPP1CC/+
F-KSame as Figure 2. G-H
Figure 2—figure supplement 2Ac306-GAL4,tsGAL80/+;UAS-NiPp1/+
Bslbo-GAL4/+;UAS-NiPp1/Pp1alpha-96A-GFP
Cw1118/Flw-YFP;slbo-GAL4/+;UAS-NiPp1/+
Figure 3A-Dc306-GAL4/+;UAS-mCherry RNAi/+
c306-GAL4/+;UAS-Pp1α−96A RNAi/+
c306-GAL4/+;UAS-Pp1-87B RNAi /+
c306-GAL4/+;UAS-Pp1-13C RNAi/+
Gc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/UAS-PLCdelta-PH-EGFP
c306-GAL4,tsGAL80/+;UAS-Pp1α−96A RNAi/+;UAS-PLCdelta-PH-EGFP/+
c306-GAL4,tsGAL80/+;UAS-Pp1-87B RNAi /+;UAS-PLCdelta-PH-EGFP/+
c306-GAL4,tsGAL80/+;UAS-Pp1-13C RNAi/+;UAS-PLCdelta-PH-EGFP/+
H-H''P{w[+mC]=Ubi mRFP.nls}1, w[*], P{ry[+t7.2]=hsFLP}12 P{ry[+t7.2]=neoFRT}19A/flwFP41 FRT 19A
Figure 3—figure supplement 1ASame as Figure 3. G
BP{w[+mC]=Ubi mRFP.nls}1, w[*], P{ry[+t7.2]=hsFLP}12 P{ry[+t7.2]=neoFRT}19A/flwFP41 FRT 19A
Figure 4A-Jc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/+
c306-GAL4,tsGAL80/+;UAS-E-cad RNAi (VDRC:103962)/+
c306-GAL4,tsGAL80/+;UAS-E-cad RNAi (VDRC:27082)/+
c306-GAL4,tsGAL80/+;UAS-β-Cat RNAi (VDRC:107344)/+
c306-GAL4,tsGAL80/+;UAS-β-Cat RNAi (VDRC:31305)/+
c306-GAL4,tsGAL80/+;UAS-α-Cat RNAi (VDRC:107298)/+
c306-GAL4,tsGAL80/+;UAS-α-Cat RNAi (VDRC:20123)/+
K-Pw1118(control)
c306-GAL4,tsGAL80/+;UAS-NiPp1/+
Figure 4—figure supplement 1A,C,E,Gc306-GAL4/+;UAS-mCherry RNAi/+
Bc306-GAL4/+;UAS-E-cad RNAi (VDRC:103962)/+
Dc306-GAL4/+;UAS-β-Cat RNAi (VDRC:107344)/+
Fc306-GAL4/+;UAS-α-Cat RNAi (VDRC:107298)/+
Gc306-GAL4/+;UAS-β-Cat RNAi (BDSC:31305)/+
I-Jc306-GAL4/+;UAS-mCherry RNAi/+
upd-GAL4,tsGAL80/+;UAS-α-Cat RNAi (VDRC:107298)/+
upd-GAL4/+;UAS-α-Cat RNAi (VDRC:20123)/+
Figure 5Ac306-GAL4,tsGAL80/+;UAS-mCherry RNAi/UAS-PLCdelta-PH-EGFP
Bc306-GAL4,tsGAL80/+;UAS-Pp1α−96A RNAi/+;UAS-PLCdelta-PH-EGFP/+
C-FSame as Figure 3. G
Figure 5—figure supplement 1B-CSame as Figure 3. G
D-Eyw; slbo-GAL4/UAS-Rac FRET (WT) and slbo-GAL4/UAS-Rac FRET; +/UAS-NiPp1
F-Jc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/UAS-PLCdelta-PH-EGFP
c306-GAL4,tsGAL80/+;UAS-α-Cat RNAi (VDRC:107298);UAS-PLCdelta-PH-EGFP/+
Figure 6Aslbo-GAL4/+;UAS-PLCdelta-PH-EGFP/+
Bslbo-GAL4/+;UAS-NiPp1/UAS-PLCdelta-PH-EGFP
D,Fw1118 (control)
E,Gslbo-GAL4/+;UAS-NiPp1/+
Hc306-GAL4,tsGAL80/+;+/sqh-GFP (VDRC:318484)
Ic306-GAL4,tsGAL80/+;UAS-NiPp1/sqh-GFP (VDRC:318484)
Figure 6—figure supplement 1ASame as Figure 6. H
BSame as Figure 6. I
Figure 6—figure supplement 2A,A',E,Ic306-GAL4,tsGAL80/+;UAS-mCherry RNAi/+
B,B',F,Jc306-GAL4,tsGAL80/+;UAS-E-cad RNAi (VDRC:103962)/+
C,C',G,Kc306-GAL4/+;UAS-β-Cat RNAi (BDSC:31305)/+
D,D',Hc306-GAL4,tsGAL80/+;UAS-sqh RNAi (VDRC:7916)/+
Figure 6—figure supplement 3A,Cc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/+
B,Dc306-GAL4,tsGAL80/+;UAS-sqh RNAi (VDRC:7916)/+
Figure 7A-A'c306-GAL4,tsGAL80/+;UAS-PLCdelta-PH-EGFP/+
B-B'c306-GAL4,tsGAL80/+;UAS-PLCdelta-PH-EGFP/UAS-NiPp1
D-Hc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/+
c306-GAL4,tsGAL80/+;UAS-Mbs RNAi/+
Figure 7—figure supplement 1D-Fw1118
G-G''FlyFos021765(pRedFlp-Hgr) (Pp1alpha-96A15346::2XTY1-SGFP-V5-preTEV-BLRP-3XFLAG)dFRT
I-I''w[1118] PBac{681 .P.FSVS-1}flw[CPTI002264]
Kc306-GAL4,tsGAL80/+;UAS-mCherry RNAi/+
Lc306-GAL4,tsGAL80/+;UAS-Mbs RNAi/+
Figure 7—figure supplement 2A-A'slbo-GAL4/UAS-Rho FRET; +/UAS-Rho FRET
B-B'slbo-GAL4/UAS-Rho FRET;UAS-NiPp1/UAS-Rho FRET
Key resources table
Reagent type
(species) or resource		DesignationSource or referenceIdentifiersAdditional
information
Genetic reagent (Drosophila melanogaster)c306-GAL4 tsGAL80Aranjuez et al., 2016Laboratory of Jocelyn McDonald
Genetic reagent (D. melanogaster)slbo-GAL4otherFBal0089668from D. Montell
Genetic reagent (D. melanogaster)upd-GAL4otherFBal0047063from D. Montell
Genetic reagent (D. melanogaster)c306-GAL4Bloomington Drosophila Stock CenterBDSC Cat# 3743; RRID:BDSC_3743
Genetic reagent (D. melanogaster)UAS-NiPp1.HABloomington Drosophila Stock CenterBDSC Cat# 23711; RRID:BDSC_23711
Genetic reagent (D. melanogaster)UAS-Pp1-87B.HABloomington Drosophila Stock CenterBDSC Cat# 24098; RRID:BDSC_24098
Genetic reagent (D. melanogaster)UAS-Pp1-13C.HABloomington Drosophila Stock CenterBDSC Cat# 23701; RRID:BDSC_23701
Genetic reagent (D. melanogaster)UAS-Pp1alpha-96A.HABloomington Drosophila Stock CenterBDSC Cat# 23700; RRID:BDSC_23700
Genetic reagent (D. melanogaster)UAS-hPPP1CCBloomington Drosophila Stock CenterBDSC Cat# 64394; RRID:BDSC_64394
Genetic reagent (D. melanogaster)UAS-mCherry RNAiBloomington Drosophila Stock CenterBDSC Cat# 35785; RRID:BDSC_35785VALIUM20-mCherry
Genetic reagent (D. melanogaster)UAS-mCD8.ChRFPBloomington Drosophila Stock CenterBDSC Cat# 27392; RRID:BDSC_27392
Genetic reagent (D. melanogaster)flwFP41 FRT 19ABloomington Drosophila Stock CenterBDSC Cat# 51338; RRID:BDSC_51338
Genetic reagent (D. melanogaster)Ubi-mRFP.nls, hsFLP, FRT19ABloomington Drosophila Stock CenterBDSC Cat# 31418; RRID:BDSC_31418
Genetic reagent (D. melanogaster)UAS-PLCδ-PH-GFPBloomington Drosophila Stock CenterBDSC Cat# 39693; RRID:BDSC_39693
Genetic reagent (D. melanogaster)UAS-Pp1α−96A RNAiVienna Drosophila Resource CenterVDRC:27673GD-11970
Genetic reagent (D. melanogaster)UAS-Pp1-87B RNAiVienna Drosophila Resource CenterVDRC:35024GD-11720
Genetic reagent (D. melanogaster)UAS-Pp1-13C RNAiVienna Drosophila Resource CenterVDRC:29057GD-14139
Genetic reagent (D. melanogaster)UAS-Mbs RNAiVienna Drosophila Resource CenterVDRC:105762KK-109231
Genetic reagent (D. melanogaster)UAS-E-cad RNAiVienna Drosophila Resource CenterVDRC:103962KK-103334
Genetic reagent (D. melanogaster)UAS-E-cad RNAiVienna Drosophila Resource CenterVDRC:27082GD-14421
Genetic reagent (D. melanogaster)UAS-β-cat RNAiVienna Drosophila Resource CenterVDRC:107344KK-102545
Genetic reagent (D. melanogaster)UAS-β-cat RNAiVienna Drosophila Resource CenterBDSC:31305TRiP.JF01252
Genetic reagent (D. melanogaster)UAS-α-cat RNAiVienna Drosophila Resource CenterVDRC:107298KK-107916
Genetic reagent (D. melanogaster)UAS-α-cat RNAiVienna Drosophila Resource CenterVDRC:20123GD-8808
Genetic reagent (D. melanogaster)fTRG sqhVienna Drosophila Resource CenterVDRC:318484fTRG 10075
Genetic reagent (D. melanogaster)fTRG Pp1α −96AVienna Drosophila Resource CenterVDRC:318084fTRG 290
Genetic reagent (D. melanogaster)flwCPTI002264Kyoto Drosophila Genomics and Genetic Resourcesline 115284FBti0143758
Genetic reagent (D. melanogaster)UAS-Flw.HAThe Zurich ORFeome Project,FlyORFline F001200
Antibodyrat monoclonal anti-E-cadherinDevelopmental Studies Hybridoma BankDCAD2; RRID:AB_5281201:10
Antibodymouse monoclonal anti-Fasciclin IIIDevelopmental Studies Hybridoma Bank7G10; RRID:AB_5282381:10
Antibodymouse monoclonal anti-ArmDevelopmental Studies Hybridoma BankN2-7A1; RRID:AB_5280891:75
Antibodymouse monoclonal anti-Fascin (Singed)Developmental Studies Hybridoma Banksn 7C; RRID:AB_5282391:25
Antibodyrabbit polyclonal anti- Phospho-Myosin Light Chain 2 (Ser19)Cell Signaling Technology, Inc#3671; RRID:AB_3302481:10
Antibodyrat monoclonal anti-HA (3F10)Millipore Sigma11867423001; RRID:AB_23146221:1000
Antibodyrabbit polyclonal anti-MbsOng et al., 20101:200 from Change Tan
Antibodyrabbit polyclonal anti-GFPThermo Fisher ScientificA11122; RRID:AB_2215691:1000–1:2000
Antibodychicken polyclonal anti-GFPAbcamab13970; RRID:AB_3007981:1000
Antibodyrabbit polyclonal anti-PPP1R8 (NiPP1)Millipore SigmaHPA027452; RRID:AB_18544901:100
AntibodyAlexa Fluor 488, 568, or 647Thermo Fisher Scientific1:400
Chemical compound, drugAlexa Fluor 488 or 568 PhalloidinThermo Fisher ScientificA12379 or A123801:400
Chemical compound, drugPhalloidin-Atto 647NMillipore Sigma659061:400
Chemical compound, drug4’,6-Diamidino-2-phenylindole (DAPI)Millipore SigmaD95420.05 µg/ml
Software, algorithmFIJIPMID:22743772
Software, algorithmGraphpad Prism 7, Prism 8https://www.graphpad.com/
Software, algorithmAdobe Photoshop CChttps://www.adobe.com/
Software, algorithmAdobe Illustrator CC 2018https://www.adobe.com/
Software, algorithmAffinity Designer 1.7.1https://affinity.serif.com/
Software, algorithmZeiss AxioVision 4.8Zeiss
Software, algorithmZeiss ZEN 3.0Zeiss
Software, algorithmFinal Cut Pro X 10.4.8Apple

Additional files

Transparent reporting form
https://cdn.elifesciences.org/articles/52979/elife-52979-transrepform-v1.pdf

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