Btlgal4

Details for all strain genotypes can be obtained from FlyBase (http://flybase.org) or in references listed here. Conditional activation of either RNAi or gene expression was achieved using the Gal4/Gal80^ts System (McGuire et al., 2004 (link)). To overexpress UAS transgenes either in all tracheal cells or in SB cells, btlGal4UASGFP;tubGal80^ts or CiGal4;tubGal80^ts was used, respectively. Crosses were kept at 18°C until late in L2 when larvae were shifted to 29°C for 48 h and dissected. The following stock flies where obtained from the Bloomington Stock Center: btlGal4 (#8807), tubGal80^ts (#7016), UASpnt^RNAi HMS01452 (#35038), UASct^RNAi JF03304 (#29625), UASEGFR^RNAi JF01368 (#25781), fzr-lacZ^G0326 (#12241), and pnt¹²⁷⁷(pntP2-lacZ, #837). UAS-fzr^RNAi (#2550 and #25553), UASbnl^RNAi (#101377 and #5730), and UASpnt^RNAi (#105390) were from Vienna Drosophila Resource Center (VDRC) and CiGal4 (Croker et al., 2006 (link)), UASbtl (Myat et al., 2005 (link)), UASbtl^DN (Reichman-Fried and Shilo, 1995 (link)), bnlGal4 (Hayashi et al., 2002 (link)), UASTor-btl (Vincent et al., 1998 (link)), UASbnl (Sutherland et al., 1996 (link)), btl-mRFPmoe (Ribeiro et al., 2004 (link)), and UASct (Ludlow et al., 1996 (link)) were given.

Fly stocks were maintained on standard cornmeal-yeast-agar medium at 25°C, and on a 12/12 hr light/dark cycle. The MiMIC and CRIMIC flies were created in the Bellen lab (see Flypush or Supplementary file 2). UAS-2xEGFP, hs-Cre,vas-dϕC31, Trojan T2A-GAL4 triplet flies were from Dr. Ben White (Diao et al., 2015 (link)). The RMCE conversion of MiMICs with GFSTF and T2A-GAL4 cassettes was described in previous studies (Diao et al., 2015 (link); Nagarkar-Jaiswal et al., 2015a (link); Nagarkar-Jaiswal et al., 2015b (link)). The crossing schemes for CRIMICs are shown in Supplemental Information 1. btl-GAL4, Ilp2-GAL4, repo-GAL4, gcm-GAL4, UAS-mCD8::GFP, UAS-mCD8::RFP, P[acman] flies, and UAS-FLP flies were obtained from the Bloomington Drosophila Stock Center (BDSC, USA). UAS-if was from Dr. Celeste Berg (Beumer et al., 1999 (link)). NP1243-GAL4, NP2222-GAL4, and NP6250-GAL4 are from Kyoto Stock Center (Kyoto DGGR, Japan). Dfs flies were from BDSC or Kyoto DGGR. UAS-cDNA flies were from BDSC or FlyORF (Switzerland). y,w;attP40(y+){nos-Cas9(v+)}/CyO (Kondo and Ueda, 2013 (link)) and y,w;+/+; attP2(y+){nos-Cas9(v+)} (Ren et al., 2013 (link)) were isogenized in this work. See Supplementary file 2 for the genotypes and stock numbers of fly stocks. All references to FlyBase are based on FlyBase 2.0/FB2017_06 (Gramates et al., 2017 (link)).