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2 protocols using uas p38a rnai

1

Drosophila Genetic Manipulation Protocols

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All crosses and staging were performed at 25°C unless otherwise noted. w1118 was used as wild-type. Stocks are described in FlyBase (http://flybase.org/). pucE69, UAS-P35 (B#5073), UAS-Scrib-RNAi (B#29552), UAS-Bsk-RNAi (B#32977), UAS-Moesin-myc (B#8631), UAS-Moesin-RNAi (B#8629), UAS-Mekk1-RNAi (B#28587), UAS-Ask1-RNAi (B#32464), UAS-Hep-RNAi (B#28710), UAS-Mkk4-RNAi (B#35140), UAS-p38b-RNAi (B#35252), UAS-p38a-RNAi (B#34744), bsk1 (B#3088), UAS-Baz-RNAi (B#39072), UAS-aPKC-RNAi (B#25946) and “Dlg-RNAi #2” (B#35286) were provided by the Bloomington Drosophila Stock Center (BDSC); Zip1 by T. Wolff (NIH Janelia Research Campus); patched-GAL4 by R. Cagan (Mount Sinai School of Medicine, New York); UAS-Cdc42- RNAi and UAS-Rho1-RNAi were described previously [11 (link),12 (link)]; “UAS-Dlg-RNAi #1”(V#41134), UAS-Rok-RNAi (V#3793), and UAS-Wnd-RNAi (V#103410) were provided by the Vienna Drosophila RNAi Center (VDRC); UAS-Slpr-RNAi, UAS-Wengen-RNAi, and UAS-Tak1-RNAi were provided by R. Fehon (University of Chicago); UAS-Egr was provided by M. Vidal (Beatson Institute, Glasgow, UK). MARCM clones were generated by heat shock at 37C for 1h and dissected 40 to 48 hours after clone induction (ACI).
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2

Genetic Manipulation of Drosophila Tissues

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Stocks were raised on standard Drosophila media, and crosses were performed at 25°C. The following stocks were described previously: w111818; TRE‐RFP19; GMR‐Gal4, ey‐Gal4, pnr‐Gal4, ptc‐Gal4, sd‐Gal4, UAS‐GFP, UAS‐Hep20; hh‐Gal421; sev‐Gal4, UAS‐dTAK122; UAS‐Wnd23; UAS‐GFP‐RNAi, UAS‐Puc, UAS‐BskDN24; UAS‐Lic, UAS‐LicKD25; UAS‐Egr26; pucE6927; and UASscrib‐RNAi, UAS‐p35.11hs‐Gal4 (#1799), UAS‐LacZ (#3956), UASp38b‐RNAi (#29405), UAS‐lic‐RNAi (#31643), UASp38a‐RNAi (#27316), UASp38c‐RNAi (#64846), UAS‐mkk4‐RNAi (#42832) and UAS‐Bsk (#9310) were obtained from the Bloomington Drosophila Stock Center. UAS‐lic‐RNAi (#20166), UAS‐wnd‐RNAi (#13786)28 and UAS‐hep‐RNAi (#26929)27 were received from the Vienna Drosophila RNAi Center. UAS‐dTAK1‐RNAi (#1388R‐2) and UAS‐p38b‐RNAi (#7393R‐1) were acquired from the National Institute of Genetics (NIG), Japan. tub‐Gal80, hs‐Flp, FRT19A; act‐Gal, UAS‐GFP (FRT19A tester) was a gift from Prof. Lei Zhang. FRT19A, licd13 mutant fly stock was a gift from Prof. Haiyun Song and has been previously described.29lic mutant clones were generated with the MARCM system 30 and labelled by GFP expression (licd13, FRT19A/tub‐Gal80, hs‐Flp, FRT19A; act‐Gal4, UAS‐GFP/+). Flp recombinase was expressed conditionally using hs‐Flp. Heat shocks were performed at 37°C for 15 minutes during the first or second instar larval stage.
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