Trh gal4

The following stocks were employed - y[1] w[*]; Mi{y[+mDint2]=MIC}Syt1[MI02197] (BDSC#35973) (Venken et al., 2011 (link)), y(1) w(*) Mi(y[+mDint2]=MIC) Sh(MI10885) (BDSC#56260), y(1) w(*);Mi(y[+mDint2]=MIC)Shal(MI10881) (BDSC#56089) (Venken et al., 2011 (link)), y(1) w(*); Mi(y[+mDint2]=MIC) Shab(MI00848) (BDSC#34115) (Venken et al., 2011 (link)), nSyb-GAL4(GMR57C10)(BDSC#39171) (Pfeiffer et al., 2008 (link)), repo-GAL4 (BDSC#7415) (Sepp et al., 2001 (link)), repo-QF2 (BDSC#66477) (Lin and Potter, 2016 (link)), Shaw-GAL4 (BDSC#60325) (Venken et al., 2011 (link); Li-Kroeger et al., 2018 (link)), Ddc-GAL4(BDSC#7009) (Feany and Bender, 2000 (link)), TH-GAL4(BDSC#8848) (Friggi-Grelin et al., 2003b (link)), Trh-GAL4(BDSC#38389) (Alekseyenko et al., 2010 (link)), UAS_H2A-GFP (Sherer et al., 2020 (link)), QUAS_H2B-mCherry (Sherer et al., 2020 (link)), brp-GAL4 (this manuscript), UAS_H2A::GFP-T2A-mKok::Caax (this manuscript). All lines were raised on standard media at 25°C, 50% RH.

Stocks were maintained on standard agar-dextrose-yeast medium at 25°C on a 12-h light/dark cycle. The Drosophila ortholog of human NGLY1 is Pngl, referred to here as dNGLY1. dNGLY1^pl flies carry an early stop codon in the dNGLY1 gene, and were previously characterized and generously provided by Perlara, PBC [20 (link)]. The dNGLY1^pl allele is homozygous lethal and the stock is maintained with the CyO balancer. The UAS-dNGLY1^wt and UAS-dNGLY1^C303A constructs were provided by Hamed Jafar-Nejad (Baylor College of Medicine) and described previously [28 (link)]. The dNGLY1^ΔGAL4 line was provided by Hugo Bellen (Baylor College of Medicine). The followings stocks were obtained from the Bloomington Drosophila Stock Center (Bloomington, IN): Trh-GAL4 (BDSC # 38389), Tubulin-GAL4 (BDSC # 5138), Dopa Decarboxylase-GAL4 (BDSC # 7009), Tyrosine Hydroxylase-GAL4 (BDSC # 8848), UAS-dNGLY1-RNAi (BDSC # 54853), UAS-sgg-RNAi (BDSC # 31308), and attp2 (RNAi control, BDSC # 36303).

Prior to experiments fly stocks were maintained at 23 °C and a 12:12 hour light [L]: dark [D] cycle on a standard yeast-sucrose-cornmeal diet. We used the following fly stocks in our experiments (BDSC = Bloomington Drosophila Stock Center; VDRC = Vienna Drosophila RNAi center): (A) GAL4 driver lines: Aug21-GAL4 (BDSC #30137); Cg-GAL4 (=collagen type IV (Col4a1) = (Cg)-GAL4; BDSC #7011); dilp2(p)-GAL4 (courtesy of Eric J. Rulifson); dilp2-GAL4 (courtesy of Linda Partridge); Gad1-GAL4 (courtesy of Pierre Leopold); hmgcr^Di-11-GAL4 (=Di-11-GAL4; courtesy of Jean-René Martin); ppl-GAL4 (courtesy of Michael J. Pankratz); Tdc2-GAL4 (BDSC #9313); TH-GAL4 (BDSC #8848); Trh-GAL4 (BDSC #38389); (B) UAS responder lines: UAS-D2R-RNAi (VDRC #11471); UAS-DopR1-RNAi (VDRC #107058); UAS-GBR-RNAi (VDRC #1784); UAS-HA-Pka-R1.BDK33 (=UAS-Pka-R33; courtesy of Franḉois Rouyer); UAS-5-HT1A-RNAi (VDRC #106094); UAS-NaChBac (courtesy of Michael O’Connor); UAS-OAMB-RNAi (VDRC #2861); UAS-Pka-R1.BDK35 (=UAS-Pka-R35, BDSC #35550); UAS-Upd2 (courtesy of Norbert Perrimon); (C) Mutant lines: Ddc^DE1 (=Ddc^hyp, BDSC #3168); dumb³ (=DopR1 hypomorph = DopR1^hyp, BDSC #19491); e¹ (BDSC #1658); ple⁴ (BDSC #3279); w¹¹¹⁸(ls-tim); and w¹¹¹⁸(s-tim) (courtesy of Charlotte Helfrich-Föster).