REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit Polyclonal anti-Gβ13F (aa 1-15)	This paper	N/A
Rabbit Polyclonal anti-Gβ13F (aa 218-233)	This paper	N/A
Mouse Monoclonal anti-β catenin	DSHB	RRID: AB_528089
Mouse Monoclonal anti-Neurotactin	DSHB	RRID: AB_528404
Chicken Polyclonal anti-GFP	Aves Labs	GFP-1020; RRID: AB_10000240
Chemicals, Peptides, and Recombinant Proteins
Colcemid-CAS 477-30-5-Calbiochem	Merck	234109-M
Gβ13F peptide (1-15aa): MNELDSLRQEAESLK	Eurogentec, This paper	UniProtKB: P26308
Gβ13F peptide (218-233aa): CKQTFPGHESDINAVT	Eurogentec, This paper	UniProtKB: P26308
Critical Commercial Assays
Direct-zol RNA MiniPrep	Zymo Research	R2050
Superscript IV VILO	Invitrogen	11766050
TaqMan Fast Advanced Master Mix	ThermoFisher	4444557
Experimental Models: Organisms/Strains
D.melanogaster: y[∗] w[67c23]	Kyoto stock center DGGR	DGRC: 101079
		Flybase: FBst0300120
D.melanogaster: P{mata4-GAL-VP16}67	Gift from E. Wieschaus	Flybase: FBti0016178
D.melanogaster: Ubi-GFP::AniRBD	This laboratory	[19]
D.melanogaster: RhoGEF2-GFP::RhoGEF2 (vk33)	Gift from Adam Martin	[24]
D.melanogaster: Sqh-Sqh::mCh	Gift from Adam Martin (constructs on chromosomes 2 and 3 have been used in all experiments at the exception of Figures 4G–4I); Another construct on chromosome 2 has been generated in this laboratory (only used in Figures 4G–4I)	[70, 71]
D.melanogaster: Sqh-eGFP:: Dp114 RhoGEF	This paper	N/A
D.melanogaster: Sqh- Dp114 RhoGEF	This paper	N/A
D.melanogaster: E-cad::GFP,sqh-Sqh::mCherry	Sqh-mCherry Gift from Adam Martin	Recombinant used in [29]
D.melanogaster: y1 sc v1; UAS- Dp114 RhoGEF shRNA	Bloomington Drosophila stock center	BL-41579
D.melanogaster: SqhPa- Dp114 RhoGEF shRNAR	This paper	N/A
D.melanogaster: SqhPa-eGFP:: Dp114 RhoGEF shRNAR	This paper	N/A
D.melanogaster: w[∗]; P{w[+mC] = mChFP Rho1}21/P{w[+mC] = mChFP-Rho1}21;	Bloomington Drosophila stock center	BL-52281 [72],
D.melanogaster: UASt-Gγ1#15	Gift from Fumio Matsuzaki	[73]
D.melanogaster: UASt-Gβ13F#20	Gift from Fumio Matsuzaki	[73]
D.melanogaster: w;FRTG13 Gγ1N159/Cyo, ftz LacZ	Gift from Fumio Matsuzaki	[73, 74]
D.melanogaster: P{w[+mW.hs] = FRT(w[hs])}G13 P{w[+mC] = ovoD1-18}2R/Dp(?;2)bw[D], S [1] wg[Sp-1] Ms(2)M [1] bw[D]/CyO	Bloomington Drosophila stock center	BL-2125
D.melanogaster: P{mata4-GAL-VP16}15	Bloomington Drosophila stock center	FBti0016179
		BL-80361
D.melanogaster: y1 sc v1;;UAS-RhoGEF2 shRNA	Bloomington Drosophila stock center	BL-34643
D.melanogaster: ;{mata4-GAL-VP16},Sqh-GFP::utABD,Sqh-Sqh::mKate;	This paper	N/A
D.melanogaster: UASt-CtaQ303L/TM3	Gift from Naoyuki Fuse	[73]
D.melanogaster: hsflp22; FRTG13 RhoGEF2l(2)04291/Cyo;	Combinant from this laboratory, FRTG13 RhoGEF2l(2)04291 is a Gift from U. Häcker	[37, 39]
D.melanogaster: UASp-GAP43::mCherry, nanos-Gal4/TM6Tb	Gift from Manos Mavrakis	N/A
D.melanogaster: y1 sc v1; UAS-Yellow shRNA /CyO	Bloomington Drosophila stock center	BL-64527
D.melanogaster: endo-α-Catenin::YFP	Cambridge Protein Trap Insertion line	CPTI-002516
		DGRC: 115551
D.melanogaster: y1 v1;UAS-Cta shRNA	Bloomington Drosophila stock center	BL-51848
D.melanogaster: w;SP/Cyo;UASp-RFP-RhoGEF2[wt]/TM3, Sb hb-lacZ{ry+}	Gift from Jörg Großhans	[75]
D.melanogaster: (y),w; 67-Gal4, UAS-EB1::GFP/CyO, ftz-lacZ	Gift from Damian Brunner	[76]
Recombinant DNA
Plasmid DNA-Casper SqhPa- Dp114RhoGEF	This paper	N/A
Plasmid DNA-Casper SqhPa- mEGFP::Dp114RhoGEF	This paper	N/A
Plasmid DNA-Casper SqhPa- Dp114RhoGEF:: mEGFP	This paper	N/A
Plasmid DNA-Casper SqhPa- Dp114RhoGEF-shRNAR	This paper	N/A
Plasmid DNA-Casper SqhPa- mEGFP::Dp114RhoGEF-shRNAR	This paper	N/A
Software and Algorithms
FlyBase	[42]	https://flybase.org/
ImageJ 1.x	[77]	https://imagej.net/Downloads
Fiji	[78]	https://imagej.net/Fiji/Downloads
Tissue Analyzer	[79]	https://grr.gred-clermont.fr/labmirouse/software/WebPA/
Stack Focuser	Michael Umorin	https://imagej.nih.gov/ij/plugins/stack-focuser.html
Axio-Vision software	Zeiss	https://www.zeiss.com/microscopy/int/products/microscope-software/axiovision.html
Zen software	Zeiss	https://www.zeiss.com/microscopy/int/products/microscope-software/zen.html
MetaMorph microscope automation and Imageanalysis software	Molecular Devices	https://www.moleculardevices.com/products/cellular-imaging-systems/acquisition-and-analysis-software/metamorph-microscopy
OriginPro9.0.0	OriginLab Corporation, Northampton, MA 01060 USA	https://www.originlab.com/