Uas rfp

Drosophila melanogaster strains were raised on standard cornmeal/agar medium supplemented with dry yeast at 25 °C with a 12 hr light/dark cycle. The wild type stock was a w¹¹¹⁸strain. The following stocks were obtained from Bloomington Stock Center: w¹¹¹⁸;dTrpA1¹ (BL26504), dTrpA1 RNAi1 (BL31384), w*;dTrpA1¹, dTrpA1-Gal4 (BL36922), w*; dTrpA1-Gal4 (BL 27593), UAS-RFP (BL 27391), UAS-GCaMP5 (BL 42037), UAS-GCaMP6m (BL 42748). The Gr66a-IRES-GFP vector was kindly provided by Kristin Scott, the NSyB-Gal4;UAS-GCaMP3 was obtained by Patrick Verstreken. The lines Gr66aGal4, UAS-dTrpA1(A);dTrpA1^ins,dTrpA1GAL4, UAS-dTrpA1(B);dTrpA1^ins,dTrpA1Gal4 and the dTrpA1 RNAi 2 were kindly provided by the laboratory of Paul Garrity.

For the transgenic RNAi and overexpression studies, the UAS-Gal4 system was employed. Flies were kept at 25°C or 29°C on standard food. RNAi stocks were obtained from the Vienna Drosophila Resource Center (Vienna, Austria): control RNAi targeting orco (100825), rap1 RNAi (20761), rap1 RNAi (110757) and magi RNAi (41735). UAS-Rap1^V12 (constitutively active), UAS-Rap1^N17 (dominant-negative), and UAS-Rap1 (wild-type) were a gift from Ulrike Gaul (LMU, Munich, Germany). The overexpression control UAS RFP (30556) was obtained from the Bloomington Stock Center (Bloomington, United States). UAS Sns and Sns-Gal4 driver lines were provided from Tobias Huber (UKE, Hamburg, Germany). For the rescue experiments, driver lines were recombined with the specific RNAi fly lines.

OR-mCD8::GFP [9 (link)], OR-Syt::GFP, OR-GAL4 [11 (link)], IR-GAL4 [52 (link)], GR-GAL4 [53 (link)] lines were from Leslie Vosshall, Barry Dickson, Richard Benton and John Carlson, respectively. Dpr-GAL4 and DIP-GAL4 [35 (link),37 (link)] lines were from Larry Zipursky. UAS-mCD8::GFP, UAS>STOP>mCD8::GFP (BL# 30125 and 30032) [54 (link)], UAS-Syt::GFP, ey-FLP, MZ19-mCD8::GFP [22 (link)], UAS-RFP, UAS-DenMark::RFP [39 (link)] (BL# 33062) UAS-dpr1, UAS-DIPγ, UAS-DIPδ, dpr10^MI03557[38 (link)] and UAS-RNAi lines were all from Bloomington Stock Center.

The primary wild-type (WT) strain was Canton S (CS), which was used for all statistics and images, except for Rhodamine 123 staining, where a second WT strain, Oregon-R (OR), was used. Two Abl alleles, Abl¹ and Abl⁴, were kept in marked and balanced stocks: Abl¹ kar red e/TM6B, Tb (from Bloomington Stock Center, Bloomington, IN) and Abl⁴ kar red e/TM6B, Tb (from Dr.FM Hoffmann), and were used for making the appropriate crosses as specified in our culture studies. The balancer line w*; Sb¹/TM3, P {ActGFP} JMR2, Ser¹ (Bloomington Stock Center, Bloomington, IN) was used to replace unlabeled balancer TM6B to make labeled heterozygous Abl alleles to facilitate genetic background checks in culture. All the Abl alleles used for cultures are Abl⁴/TM3-ser-GFP and Abl¹//TM3-Ser-GFP. CS was crossed with both Abl⁴/TM3-ser-GFP and Abl¹//TM3-ser-GFP to yield heterozygous Abl alleles under wild-type background for cultures. The ubiquitously expressed driver, daughterless (Da-Gal4) (Gift from Toshi Kitamoto) and the pan-neuronal driver, Elav-Gal4, were used to drive UAS-GFP, UAS-RFP (Bloomington Stock Center, Bloomington, IN).