The following Drosophila stocks were used: UAS-Vha68-2RNAi (34390; Vienna Drosophila Resource Center [VDRC] and BL34582; Bloomington Drosophila Stock Center), UAS-Vha100-2RNAi (30297; VDRC), UAS-VhaAC39-1RNAi (20950; VDRC), E(spl)mγ-GFP (Almeida and Bray, 2005 (link)), NRE-LacZ; esg-GAL4, UAS-GFP, tubulin-GAL80ts (Lucchetta and Ohlstein, 2017 (link)), UAS-NotchRNAi (100002; VDRC), UAS-NotchΔECD (Larkin et al., 1996 (link)), UAS-bratRNAi (31333; VDRC), and UAS-mCherryRNAi (BL35785). The following GAL4 driver lines were used: UAS-dcr2; ase-GAL4, UAS-stingerGFP (Barolo et al., 2000 (link)), ase-GAL4 (Zhu et al., 2006 (link)), UAS-dcr2; wor-GAL4, ase-GAL80 (Neumüller et al., 2011 (link)), UAS-stingerRFP (Homem et al., 2014 (link)), and esg-GAL4, UAS-mCD8::GFP, tubulin-GAL80ts (Goulas et al., 2012 (link)).
Stock generated in this study was Vha68-2-GFP. gRNA was cloned into pU6-BbsI-chiRNA (45946; 100 ng/μl; Addgene) and coinjected with the donor plasmid (250 ng/μl) into act>Cas9 embryos. The gRNA used was GGAGGACTAGAGACCGCGC.
Fly crosses were set up at 25°C for 24 h and then shifted to 29°C. For experiments inFig. 7 (b–f) , fly crosses were set up and reared at 18°C. 3-d-old female flies were shifted to 29°C for 6 d. For the brat rescue, double RNAi crosses were set up and reared at 29°C. Flies were collected 2 d after eclosion and kept at 29°C.
Stock generated in this study was Vha68-2-GFP. gRNA was cloned into pU6-BbsI-chiRNA (45946; 100 ng/μl; Addgene) and coinjected with the donor plasmid (250 ng/μl) into act>Cas9 embryos. The gRNA used was GGAGGACTAGAGACCGCGC.
Fly crosses were set up at 25°C for 24 h and then shifted to 29°C. For experiments in
