Sev gal4

All stocks were raised on standard Drosophila media and crosses were performed at 25 °C unless otherwise indicated. For experiments involving tub-Gal80^ts, flies were raised at 18 °C to restrict Gal4 activity for 5–6 days, then shifted to 29 °C for 2 days to inactivate Gal80^ts. The following stocks were used: GMR-Gal4, ptc-Gal4, sev-Gal4, UAS-GFP, UAS-Rac1 (6680), UAS-Rho1 (7334), UAS-LacZ (3956) and wnd^Exel6135 (7614, EP line use for overexpression), all obtained from the Bloomington Stock Center (Bloomington, IN, USA), UAS-Rac1-IR (2248R-1)^{43 (link)} obtained from National Institute of Genetics (NIG, Mishima, Japan), UAS-Wnd^KD, wnd¹, wnd³ (gifts from Aaron DiAntonio, St. Louis, MO, USA), UAS-Ask1^DN (gift from Masayuki Miura, Tokyo, Japan), hep¹, UAS-Egr, UAS-dTAK1, UAS-dTAK1^DN, UAS-Bsk^DN, UAS-hep-IR, UAS-Puc, puc^E69,^{44 (link)}bsk¹,^{34 (link)}UAS-Hep^CA, dTAK1¹,^{27 (link)}UAS-wnd-IR,^{24 (link)}UAS-MKK4-IR,^{43 (link)}mkk4^G673,^{37 (link)}UAS-slpr-IR^{18 (link)} and UAS-mekk1-IR,^{45 (link)} as previously described.

Fluorescently labeled clones were produced in larval imaginal discs using the following strains: y, w, eyFLP1; Act>y+>Gal4, UAS–GFP; FRT82B, Tub-Gal80 (82B tester) and y, w, eyFLP1; Tub-Gal80, FRT40A; Act>y+>GAL4, UAS-GFP (40A tester). Additional strains used were as follows: GMR-GAL4, ptc-GAL4, ap-GAL4, sev-GAL4, UAS-GFP and puc^E69 (puc-lacZ) were obtained from Bloomington Drosophila Stock Center. UAS-spz^ACT was gift from J.-M. Reichart (Ligoxygakis et al., 2002 (link)). Five independent UAS-Toll-6-IR transgenic lines generated from three different constructs were obtained from Vienna Drosophila Resource Center (VDRC). UAS-spz5^HA was obtained from FlyORF. UAS-egr (Igaki et al., 2002 (link)), UAS-Hep^CA, UAS-dTAK1, UAS-hep-IR, UAS-bsk^DN, UAS-dTAK1^DN and UAS-puc (Ma et al., 2012 (link)) were previously described. UAS-Toll^WT and UAS-Toll-6^ACT-Flag transgenic flies were generated by standard P-element-mediated transformation (Bestgene, Inc.). More than five independent lines were produced and examined for each transgene. Two RNAi lines (v27102 and v27103) were recombined and used to perform experiments unless indicated. Gene expression was verified by immunostaining.

The flies were raised at 25°C unless otherwise mentioned.
The following flies were used in this study:
CantonS, FRT42B, FRT2A, Gal80/II, Gal80/III, FRT80, GMR-GFP, UAS-mCD8GFP, hs-FLP, ey3.5-FLP, GMR-FLP, tub-Gal80ts, elav-Gal4, sev-Gal4, LGMR-Gal4, m∂0.5-Gal4, ro-tau-LacZ, Rh6-EGFP, PanR7-Gal4 were obtained from Bloomington Drosophila Stock Center. sens-Gal4/CyO and sens-Gal4/TM6 were gift from Bassem Hassan. sequoia⁵ is a previously generated Sequoia loss-of-function allele (Petrovic and Hummel, 2008 (link)). UAS-Sequoia was obtained from Jay Brenman. UAS-Capricious, UAS-Capricious^ID(intracellular deletion), Capricious^C18fs FRT2A and caps-LacZ^nls flies were kindly provided by Akinao Nose. GMR-gogo was a generous gift from Takashi Suzuki. PanR8-Gal4 was a gift from Claude Desplan. UAS-Capricious^RNAi was obtained from VDRC (VDRC Transformant ID No. 27097). PM181-Gal4 and CadN⁴⁰⁵FRT40 (Lee et al., 2001 (link)) were used in Sequoia analysis, Early Dm8 labelling OK371-VP16AD/CyO; ortC2-Gal4DBD/TM2, adult Dm8 specific OrtC1-3 LexA DBD, OrtC2B dVP16AD/CyO (Ting et al., 2014 (link)) and syn-GRASP constructs UAS-Syb::spGFP1-10 and LexAop spGFP11::CD4/TM2 (Karuppudurai et al., 2014 (link)) were used in syb-GRASP experiments.