1. Stem Cells and Regenerative Medicine

Sequential activation of transcriptional repressors promotes progenitor commitment by silencing stem cell identity genes

  1. Noemi Rives-Quinto
  2. Hideyuki Komori
  3. Cyrina M Ostgaard
  4. Derek H Janssens
  5. Shu Kondo
  6. Qi Dai
  7. Adrian W Moore
  8. Cheng-Yu Lee  Is a corresponding author
  1. Life Sciences Institute, University of Michigan, United States
  2. Department of Cell and Developmental Biology, University of Michigan Medical School, United States
  3. Invertebrate Genetics Laboratory, National Institute of Genetics, Japan
  4. Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Sweden
  5. Riken Center for Brain Science, Japan
  6. Division of Genetic Medicine, Department of Internal Medicine and Comprehensive Cancer Center, University of Michigan Medical School, United States
https://doi.org/10.7554/eLife.56187
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Cite this article
  1. Noemi Rives-Quinto
  2. Hideyuki Komori
  3. Cyrina M Ostgaard
  4. Derek H Janssens
  5. Shu Kondo
  6. Qi Dai
  7. Adrian W Moore
  8. Cheng-Yu Lee
(2020)
Sequential activation of transcriptional repressors promotes progenitor commitment by silencing stem cell identity genes
eLife 9:e56187.
https://doi.org/10.7554/eLife.56187
  2. Download BibTeX
  3. Download .RIS
7 figures, 1 table and 1 additional file

Figures

Figure 1 with 1 supplement
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Identification of candidate regulators of type II neuroblast functional identity.

(A) A diagram of the type II neuroblast lineage showing the expression patterns of genes and Gal4 drivers used throughout this study. (B) Gene transcription profiles of brat-null brains transiently …

Figure 1—figure supplement 1
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Time-course analysis of transient Insb overexpression in brat-null brains.

(A) Quantification of total type II neuroblasts (top) or INPs (bottom) per brat-null brain lobe that transiently overexpressed Insb driven by a type II neuroblast Gal4. Insb overexpression led to …

Figure 1—figure supplement 1—source data 1

Quantification of total type II neuroblasts or INPs per brat-null brain lobe that transiently overexpressed Insb.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig1-figsupp1-data1-v2.xlsx
Figure 2
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tll is necessary and sufficient for type II neuroblast functional identity.

(A) A diagram showing the expression patterns of genes in the type I and II neuroblast lineages. (B) Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-tllRNAi …

Figure 2—source data 1

Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-tllRNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig2-data1-v2.xlsx
Figure 2—source data 2

Quantification of total type II neuroblasts per brain that overexpressed a UAS-tll transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig2-data2-v2.xlsx
Figure 2—source data 3

Quantification of total ventral type I neuroblasts per brain lobe that overexpressed a UAS-tll transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig2-data3-v2.xlsx
Figure 3 with 1 supplement
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Ham is a novel regulator of INP commitment.

(A) Quantification of total type II neuroblasts per ermhypo brain lobe that overexpressed a UAS-RNAi transgene driven by a type II neuroblast Gal4. Knocking-down ham function consistently enhanced …

Figure 3—source data 1

Quantification of total type II neuroblasts per ermhypo brain lobe that overexpressed a UAS-RNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig3-data1-v2.xlsx
Figure 3—source data 2

Quantification of total type II neuroblasts per ham-mutant brain lobe.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig3-data2-v2.xlsx
Figure 3—source data 3

Quantification of total type II neuroblasts per brain lobe of the indicated genotypes.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig3-data3-v2.xlsx
Figure 3—source data 4

Quantification of total type II neuroblasts per erm,ham double heterozygous brain lobe that overexpressed a UAS-ham transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig3-data4-v2.xlsx
Figure 3—figure supplement 1
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Ham functions synergistically with Erm to suppress supernumerary type II neuroblast formation.

(A) Quantification of total type II neuroblasts per brain lobe of the indicated genotypes. erm,ham double heterozygous brains displayed the supernumerary type II neuroblast phenotype. (B) …

Figure 3—figure supplement 1—source data 1

Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-hamRNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig3-figsupp1-data1-v2.xlsx
Figure 4
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Ham suppresses INP reversion by repressing gene transcription.

(A–B) Images of wild-type or hamSK1 homozygous type II neuroblast mosaic clones. Supernumerary neuroblasts (−15 μm) in hamSK1 homozygous clones were always located far from the parental neuroblast …

Figure 4—source data 1

Quantification of total neuroblasts per ham1 or hamSK1 homozygous type II neuroblast clone.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig4-data1-v2.xlsx
Figure 4—source data 2

Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-ham transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig4-data2-v2.xlsx
Figure 4—source data 3

Quantification of total type II neuroblasts per hamSK1 homozygous brain lobe that overexpressed various UAS-ham transgenes.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig4-data3-v2.xlsx
Figure 4—source data 4

Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-VP16::hamN-ZF transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig4-data4-v2.xlsx
Figure 5
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Erm- and Ham-mediated repression renders tll refractory for activation by Notch.

(A) A diagram depicting our hypothesis that ectopic activation of tll in INPs leads to supernumerary type II neuroblasts in erm,ham double heterozygous brains. (B) Quantification of total type II …

Figure 5—source data 1

Quantification of total type II neuroblasts per brain lobe that was erm,ham double heterozygous or erm,ham,tll triple heterozygous.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig5-data1-v2.xlsx
Figure 5—source data 2

Quantification of total type II neuroblasts erm,ham double heterozygous brain lobe that carried one copy of the tll::GFP(BAC) transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig5-data2-v2.xlsx
Figure 5—source data 3

Quantification of Tll::GFP expression relative to Dpn expression in type II neuroblasts that mis-expressed a UAS-erm or UAS-ham transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig5-data3-v2.xlsx
Figure 5—source data 4

Quantification of total type II neuroblasts per wild-type or hamSK1 homozygous brain lobe that overexpressed a UAS-NRNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig5-data4-v2.xlsx
Figure 5—source data 5

Quantification of total type II neuroblasts per erm or ham heterozygous brain lobe that overexpressed a UAS-Nintra transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig5-data5-v2.xlsx
Figure 6 with 1 supplement
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Erm and Ham function through Hdac3 to prevent INPs from reverting to type II neuroblasts.

(A) Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-ham transgene driven by an INP Gal4. Overexpressing full-length Ham or HamΔC-ZF in INPs suppressed the …

Figure 6—source data 1

Quantification of total type II neuroblasts per brain lobe that overexpressed a UAS-ham transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig6-data1-v2.xlsx
Figure 6—source data 2

Quantification of total type II neuroblasts per ham heterozygous brain lobe that overexpressed various UAS transgenes.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig6-data2-v2.xlsx
Figure 6—source data 3

Quantification of total type II neuroblasts per erm heterozygous brain lobe that overexpressed a UAS-hdac3RNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig6-data3-v2.xlsx
Figure 6—source data 4

Quantification of Tll::GFP expression relative to Dpn expression in INPs derived from type II neuroblasts that overexpressed a UAS-hdac3RNAi transgene.

https://cdn.elifesciences.org/articles/56187/elife-56187-fig6-data4-v2.xlsx
Figure 6—figure supplement 1
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Ham overexpression in Ase+ immature INPs suppressed INP reversion in erm-null brains.

(A–B) Images of erm-null brains that overexpressed a UAS-ham transgene driven by an INP Gal4. Ham overexpression in INPs suppressed the supernumerary type II neuroblast phenotype in erm-null brains. …

Figure 7
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A proposed model for the regulation of type II neuroblast functionality.

We propose that Erm and Ham recruits Hdac3 to silence tll during INP commitment, preventing re-activation of Notch signaling in INPs from triggering Tll expression in wild-type brains. The tll locus …

Tables

Appendix 1—key resources table
Reagent type
(species) or resource		DesignationSource or referenceIdentifiersAdditional information
Antibodyanti-GFP
(Chicken polyclonal)		Aves Labs, INC.Cat#GFP-1020,
RRID:AB_2307313		IF(1:2000)
Antibodyanti-V5
(Mouse monoclonal)		ThermoFisher ScientificCat#R960-25,
RRID:AB_2556564		IF(1:500)
Antibodyanti-Ase
(Rabbit polyclonal)		Weng et al., 2010 doi: 10.1016/j.devcel.2009.12.007.IF(1:400)
Antibodyanti-Hamlet (Rabbit polyclonal)Eroglu et al., 2014 doi: 10.1016/j.cell.2014.01.053.IF(1:50)
Antibodyanti-Dpn
(Rat monoclonal)		Lee et al., 2006a doi: 10.1038/nature04299.clone 11D1BC7.14IF(1:2)
AntibodyAlexa Fluor 488 AffiniPure Anti-Chicken IgY (IgG) (H+L)
(Donkey polyclonal)		Jackson Immuno
Research Laboratories, INC.		Cat#703-545-155,
RRID:AB_2340375		IF(1:500)
AntibodyAlexa Fluor 647 AffiniPure anti-Rat IgG (H+L)
(Goat polyclonal)		Jackson Immuno Research Laboratories, INC.Cat#112-605-167
RRID:AB_2338404		IF(1:500)
Antibodyanti-Mouse IgG (H+L) Highly Cross-Adsorbed
Secondary Antibody, Alexa Fluor 488
(Goat polyclonal)		ThermoFisher
Scientific		Cat#A-11029,
RRID:AB_2534088		IF(1:500)
Antibodyanti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Goat polyclonal)ThermoFisher ScientificCat#A-11034,
RRID:AB_2576217		IF(1:500)
Antibodyanti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 (Goat polyclonal)ThermoFisher ScientificCat#A-11035,
RRID:AB_2534093		IF(1:500)
OtherRhodamine PhalloidinThermoFisher ScientificCat#R415IF(1:100)
Genetic reagent (D. melanogaster)brat11/CyO, Actin-GFPLee et al., 2006b doi: 10.1016/j.devcel.2006.01.017.
Genetic reagent (D. melanogaster)w1118; Df(2L)Exel8040/CyOBloomington
Drosophila Stock Center		BDSC: 7847
FlyBase: FBst0007847;
RRID:BDSC_7847		FlyBase symbol: Df(2L)Exel8040/CyO
Genetic reagent (D. melanogaster)y1, M{vas-int.Dm}ZH-2A w*; M{UAS-insbFL-myc}ZH-86FbKomori et al., 2018 doi: 10.1101/gad.320333.118.
Genetic reagent (D. melanogaster)Wor-Gal4(II)Lee et al., 2006a doi: 10.1038/nature04299.
Genetic reagent (D. melanogaster)Wor-Gal4(III)Weng et al., 2010 doi: 10.1016/j.devcel.2009.12.007.
Genetic reagent (D. melanogaster)y1, w*; P{tubPGAL80} LL10, P{neoFRT}40A/CyOBloomington Drosophila Stock CenterBDSC: 5192
FlyBase: FBst0005192;
RRID:BDSC_5192		FlyBase symbol: y1, w*; P{tubPGAL80} LL10, P{neoFRT}40A/CyO
Genetic reagent (D. melanogaster)P{hsFLP}1, P{tubP-GAL80}LL1, w*,
P{neoFRT}19A; P{UAS-mCD8::GFP.L}LL5		Bloomington Drosophila Stock CenterBDSC: 5134
FlyBase: FBst0005134;
RRID:BDSC_5134_		FlyBase symbol: P{hsFLP}1, P{tubP-GAL80}LL1, w*,
P{neoFRT}19A; P{UAS-mCD8::GFP.L}LL5
Genetic reagent (D. melanogaster)y1 w*; PBac{y[+mDint2] w[+mC]=tll EGFP.S}VK00037Bloomington Drosophila Stock CenterBDSC: 30874
FlyBase: FBst0030874;
RRID:BDSC_30874		FlyBase symbol: y1 w*; PBac{y[+mDint2] w[+mC]=tll EGFP.S}VK00037
Genetic reagent (D. melanogaster)Erm-Gal4 (II)Pfeiffer et al., 2008 doi: 10.1073/pnas.0803697105.
Genetic reagent (D. melanogaster)Erm-Gal4 (III)Pfeiffer et al., 2008 doi: 10.1073/pnas.0803697105.
Genetic reagent (D. melanogaster)tllRNAi: y1 sc* v1 sev21; P{TRiP.HMS01316}attP2Bloomington Drosophila Stock CenterBDSC: 34329
FlyBase: FBst0034329;
RRID:BDSC_34329		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS01316}attP2
Genetic reagent (D. melanogaster)M{UAS-tll.ORF-VN}ZH-86FbFlyORFF004752 FBst0502964;
RRID:FlyORF_ F004752		FlyBase symbol: M{UAS-tll.ORF-VN}ZH-86Fb
Genetic reagent (D. melanogaster)Ase-Gal80 (II)Neumüller et al., 2011 doi: 10.1016/j.stem.2011.02.022.
Genetic reagent (D. melanogaster)erm1/CyO, Act-GFPWeng et al., 2010 doi: 10.1016/j.devcel.2009.12.007.
Genetic reagent (D. melanogaster)erm2/CyO, Act-GFPWeng et al., 2010 doi: 10.1016/j.devcel.2009.12.007.
Genetic reagent (D. melanogaster)UAS-ermWeng et al., 2010 doi: 10.1016/j.devcel.2009.12.007.
Genetic reagent (D. melanogaster)PBac{erm-flag4C(g)}VK33Janssens and Lee, 2014 doi: 10.1242/dev.106534.
Genetic reagent (D. melanogaster)DRNAi: y1 v1; P{TRiP.JF02115}attP2Bloomington Drosophila Stock CenterBDSC: 26217
FlyBase: FBst0026217;
RRID:BDSC_26217		FlyBase symbol: y1 v1; P{TRiP.JF02115}attP2
Genetic reagent (D. melanogaster)AseRNAi: y1 sc* v1 sev21; P{TRiP.HMS02847}attP2Bloomington Drosophila Stock CenterBDSC: 44552
FlyBase: FBst0044552;
RRID:BDSC_44552		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS02847}attP2
Genetic reagent (D. melanogaster)hamRNAi: y1 v1; P{TRiP.JF02270}attP2Bloomington Drosophila Stock CenterBDSC: 26728
FlyBase: FBst0026728;
RRID:BDSC_26728		FlyBase symbol: y1 v1; P{TRiP.JF02270}attP2
Genetic reagent (D. melanogaster)hamRNAi: y1 sc* v1 sev21; P{y[+t7.7] v[+t1.8]=TRiP.HMS00470}attP2Bloomington Drosophila Stock CenterBDSC: 32470
FlyBase: FBst0032470;
RRID:BDSC_32470		FlyBase symbol: y1 sc* v1 sev21; P{y[+t7.7] v[+t1.8]=TRiP.HMS00470}attP2
Genetic reagent (D. melanogaster)OpaRNAi: y1 sc* v1 sev21; P{TRiP.HMS01185}attP2/TM3, Sb1Bloomington Drosophila Stock CenterBDSC: 34706
FlyBase: FBst0034706;
RRID:BDSC_34706		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS01185}attP2/TM3, Sb1
Genetic reagent (D. melanogaster)w1118; Df(2L)Exel7071/CyOBloomington Drosophila Stock CenterBDSC: 7843
FlyBase: FBst0007843;
RRID:BDSC_7843		FlyBase symbol: w1118; Df(2L)Exel7071/CyO
Genetic reagent (D. melanogaster)hamSKI, FRT40A/CyOThis paperA new hamlet mutant fly line
Genetic reagent (D. melanogaster)P{w[+mW.hs]=GawB}elav[C155],
P{w[+mC]=UAS-mCD8::GFP.L}Ptp4E[LL4],
P{ry[+t7.2]=hsFLP}1, w[*]		Bloomington Drosophila Stock CenterBDSC: 5146
FlyBase: FBst0005146;
RRID:BDSC_5146		FlyBase symbol: P{w[+mW.hs]=GawB}elav[C155],
P{w[+mC]=UAS-mCD8::GFP.L}Ptp4E[LL4],
P{ry[+t7.2]=hsFLP}1, w[*]
Genetic reagent (D. melanogaster)w1118; P{w[+mC]=UAS-Dcr-2.D}2Bloomington Drosophila Stock CenterBDSC: 24650
FlyBase: FBst00024650;
RRID:BDSC_24650		FlyBase symbol: w1118; P{w[+mC]=UAS-Dcr-2.D}2
Genetic reagent (D. melanogaster)w*; P{w[+mC]=tubP- GAL8ts}2/TM2Bloomington Drosophila Stock CenterBDSC: 7017
FlyBase: FBst00024650;
RRID:BDSC_7017		FlyBase symbol: w*; P{w[+mC]=tubP- GAL8ts}2/TM2
Genetic reagent (D. melanogaster)ham1,FRT40A/CyoMoore et al., 2002 doi: 10.1126/science.1072387.
Genetic reagent (D. melanogaster)UAS-hamMoore et al., 2002 doi: 10.1126/science.1072387.
Genetic reagent (D. melanogaster)RNAi of Notch: y1, v1; P{y+t7.7v+t1.8=TRiP.HMS00001}attP2Bloomington Drosophila Stock CenterBDSC: 33611
FlyBase: FBst0033611;
RRID:BDSC_33611		FlyBase symbol: y1, v1; P{y+t7.7v+t1.8=TRiP.HMS00001}attP2
Genetic reagent (D. melanogaster)Oregon-R-CBloomington Drosophila Stock CenterBDSC: 5
FlyBase: FBst0000005;
RRID:BDSC_5		FlyBase symbol: Oregon-R-C
Genetic reagent (D. melanogaster)P{hsFLP}1, y1 w*; P{UAS- N.intra.GS}2/CyO; MKRS/TM2Bloomington Drosophila Stock CenterBDSC: 52008
FlyBase: FBst0052008;
RRID:BDSC_52008		FlyBase symbol: P{hsFLP}1, y1 w*; P{UAS- N.intra.GS}2/CyO; MKRS/TM2
Genetic reagent (D. melanogaster)y1; M{vas-int.Dm}ZH-2A w*;
M{UAS-ham∆C-ZF-myc}ZH-86Fb		This paperTransgene expressing Hamlet mutant form of the C-terminal zinc finger deletion version
Genetic reagent (D. melanogaster)y1; M{vas-int.Dm}ZH-2A w*; M{UAS-ERD::hamN-ZF-myc}ZH-86FbThis paperTransgene expressing Hamlet the N-terminal zinc finger fused with ERD transcriptional repression domain
Genetic reagent (D. melanogaster)y1; M{vas-int.Dm}ZH-2A w*;
M{UAS-VP-16::hamN-ZF-myc}ZH-86Fb		This paperTransgene expressing Hamlet the N-terminal zinc finger fused with VP16 transcriptional activatoin domain
Genetic reagent (D. melanogaster)cu1, tll49/TM3, P{ftz/lacC}SC1, Sb1, Ser1Bloomington Drosophila Stock CenterBDSC: 7093
FlyBase: FBst007093;
RRID:BDSC_7093		FlyBase symbol: cu1, tll49/TM3, P{ftz/lacC}SC1, Sb1, Ser1
Genetic reagent (D. melanogaster)st1 e1 tll1/TM3, Sb1Bloomington Drosophila Stock CenterBDSC: 2729
FlyBase: FBst002729;
RRID:BDSC_2729		FlyBase symbol: st1 e1 tll1/TM3, Sb1
Genetic reagent (D. melanogaster)hamSK4, FRT40A/CyOBloomington Drosophila Stock CenterBDSC: 34329FlyBase: FBst0034329;RRID:BDSC_34329FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS01316}attP2
Genetic reagent (D. melanogaster)hdac3RNAi: y1 sc* v1 sev21; P{TRiP.HMS00087}attP2Bloomington Drosophila Stock CenterBDSC: 34778
FlyBase: FBst0034778;
RRID:BDSC_34778		FlyBase symbol: hdac3RNAi: y1 sc* v1sev21; P{TRiP.HMS00087}attP2
Genetic reagent (D. melanogaster)Su(z)12RNAi: y1 sc* v1 sev21; P{TRiP.HMS00280}attP2/TM3, Sb1Bloomington Drosophila Stock CenterBDSC: 33402
FlyBase: FBst0033402;
RRID:BDSC_33402		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS00280}attP2/TM3, Sb1
Genetic reagent (D. melanogaster)Su(var)3-3RNAi: y1 sc* v1 sev21; P{TRiP.HMS00638}attP2Bloomington Drosophila Stock CenterBDSC: 32853
FlyBase: FBst0032853;
RRID:BDSC_32853		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.HMS00638}attP2
Genetic reagent (D. melanogaster)Su(var)205RNAi: y1 sc* v1 sev21; P{TRiP.GL00531}attP40Bloomington Drosophila Stock CenterBDSC: 36792
FlyBase: FBti0146447;
RRID:BDSC_36792		FlyBase symbol: y1 sc* v1 sev21; P{TRiP.GL00531}attP40
Genetic reagent (D. melanogaster)UAS-Mi-2DNKovač et al., 2018 doi: 10.1038/s41467-018-04503-2.
Genetic reagent (D. melanogaster)UAS-Mi-brmDNHerr et al., 2010 doi: 10.1016/j.ydbio.2010.04.006.
Genetic reagent (D. melanogaster)In(1)wm4; Su(var)3–91/TM3, Sb1 Ser1Bloomington Drosophila Stock CenterBDSC: 6209
FlyBase: FBst0006209;
RRID:BDSC_6209		FlyBase symbol: In(1)wm4; Su(var)3–91/TM3, Sb1 Ser1
Genetic reagent (D. melanogaster)w1118; PBac{Sp1- EGFP.S}VK00033Bloomington Drosophila Stock CenterBDSC: 38669
FlyBase: FBst0038669;
RRID:BDSC_38669		FlyBase symbol: w1118; PBac{Sp1- EGFP.S}VK00033
Sequenced-based reagentHam_FEroglu et al., 2014 doi: 10.1016/j.cell.2014.01.053.PCR primersatagatcctttggccagcagac
Sequenced-based reagentHam_REroglu et al., 2014 doi: 10.1016/j.cell.2014.01.053.PCR primersagtactcctccctttcggcaat
Sequenced-based reagentAse_FKomori et al., 2014b doi: 10.7554/eLife.03502.PCR primersagcccgtgagcttctacgac
Sequenced-based reagentAse_RKomori et al., 2014b doi: 10.7554/eLife.03502.PCR primersgcatcgatcatgctctcgtc
Sequenced-based reagentD_FThis paperPCR primersgcggcggcggtcaacaat
Sequenced-based reagentD_RThis paperPCR primerstgcggcgtacagcgaagggt
Sequenced-based reagentErm_FEroglu et al., 2014 doi: 10.1016/j.cell.2014.01.053.PCR primersgttacggccaggcatcgggtcaa
Sequenced-based reagentErm_REroglu et al., 2014 doi: 10.1016/j.cell.2014.01.053.PCR primersgggccaggcgggattactcgtctc
Sequenced-based reagentPntP1_FKomori et al., 2014b doi: 10.7554/eLife.03502.PCR primersggcagtacgggcagcaccac
Sequenced-based reagentPntP1_RKomori et al., 2014b doi: 10.7554/eLife.03502.PCR primersctcaacgcccccaccagatt
Sequenced-based reagentDpn_FKomori et al., 2014b doi: 10.7554/eLife.03502.Komori et al., 2014bPCR primerscatcatgccgaacacaggtt
Sequenced-based reagentDpn_RKomori et al., 2014bPCR primersgaagattggccggaactgag
Recombinant DNA reagentpUAST-ham∆C-ZF-myc-attB (plasmid)This paperPlasmid DNA of a transgene expressing Hamlet mutant form of the C-terminal zinc finger deletion version
Recombinant DNA reagentpUAST-ERD::hamN-ZF-myc-attB (plasmid)This paperPlasmid DNA of a transgene expressing Hamlet the N-terminal zinc finger fused with ERD transcriptional repression domain
Recombinant DNA reagentpUAST-VP16::hamN-ZF-myc-attB (plasmid)This paperPlasmid DNA of a transgene expressing Hamlet the N-terminal zinc finger fused with VP16 transcriptional activatoin domain
Software, algorithmLAS AFLeica MicrosystemsRRID:SCR_013673
Software, algorithmImageJ 1.50 gNational Institute of HealthRRID:SCR_003070

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