Uas pka c1

Fly stocks were maintained at room temperature using standard techniques. Experimental crosses were raised at 25°C in 60% relative humidity. The following flies were used in this study: UAS-HDAC4 (a gift from Biao Wang); tdc2^R054 and tbh^nM18 are gifts from Vivian Budnick; dVMAT^P1 (a gift from David Krantz); puc-lacZ; UAS-mito-GFP; eag¹ shaker¹²⁰ (a gift from Troy Littleton). Flies obtained from the Bloomington Drosophila Stock Center include Repo-GAL4, Nrv2-Gal4, UAS-LexA RNAi (RRID:BDSC_67945), UAS-PKA-C1 (RRID:BDSC_35554), UAS-PKA-R1 RNAi (RRID:BDSC_27308), UAS-G_sα. Q215L (RRID:BDSC_6490), UAS-nls-GFP (RRID:BDSC_4775), UAS-HDAC4 RNAi (RRID:BDSC_28549, BDSC_34774), UAS-Octβ1R RNAi (RRID:BDSC_31107), and UAS-OAMB RNAi (RRID:BDSC_31171). UAS-SIK3 RNAi (KK 106268) flies were obtained from the Vienna Drosophila Resource Center. In GAL4 experiments, unless otherwise noted, control flies were generated by crossing virgin females with the GAL4 driver to UAS-LexA RNAi male flies.

CantonS, elav-GAL4, UAS-PKA-C1, UAS-DsRed, GMR52D11-lexA and P{UAS-CD4-spGFP1–10}3, P{lexAop-CD4-spGFP11}3 were obtained from the Bloomington Stock Center. Pka-C3^NIG.6117R was obtained from the National Institute of Genetics (NIG-Fly), Japan. natalisin-GAL4 was kindly provided by Y. Park and Y-J. Kim and is described in^{25 (link)}. PBacf07226 and PBacPka-C3^f00695 were obtained from the Exelixis Collection at the Harvard Medical School. UAS-PKA-C3 is described in^{21 (link)}. The constitutively inactive PKA-C3 construct was generated by replacing aspartate 397 (D³⁹⁷, see Supp. Figure 3) with alanine by site–directed mutagenesis using the site-directed mutagenesis kit from clontech. D³⁹⁷ is part of the highly conserved YRDLKPEN core sequence of the catalytic loop and mutations in the conserved aspartate have been shown to abolish or dramatically reduce the catalytic activity^{36 (link)–38 (link)}. The construct was tagged with HA. Flies were maintained on standard fly food under a 12:12 h light:dark cycle. Stocks were maintained at 18 °C while crosses and aging flies were maintained at 25 °C. The age of the flies in each experiment is indicated in the figures and figure legends.