Flies were raised on cornmeal agar food at 24°, 70% humidity, and with a 14:10 light:dark cycle. All flies used in this study were male. Male 4- to 5-day-old flies were isolated under CO2, given 1 day to recover, and then conditioned with specified paradigms detailed below. The fly lines used in this study include two pan-MB specific drivers, a MB010B split Gal4 line [#68293; Bloomington Drosophila Stock Center (BDSC)] (Aso et al. 2014) and R19B03 Gal4 line (#49830; BDSC) (Jenett et al. 2012), tubP-Gal80ts-7 (#7018; BDSC) (McGuire et al. 2004) recombined with R19B03-Gal4, a 5xUAS-unc84-2xGFP “INTACT” line (Henry et al. 2012), UAS-Cdc5-RNAi (#57425; BDSC) (Perkins et al. 2015), UAS-Rm62-RNAi (#34829; BDSC) (Perkins et al. 2015), UAS-Ref1-RNAi (#34626; BDSC) (Perkins et al. 2015), UAS-Pep-RNAi (#32944; BDSC) (Perkins et al. 2015), UAS-CG7971-RNAi (#52936; BDSC) (Perkins et al. 2015), GFP-RNAi (9330; BDSC), yw (Janelia Research Campus), and TRiP control (#36303; BDSC) (Perkins et al. 2015). elavC155-Gal4 (#458; BDSC) and Act5C-Gal4 (#25374; BDSC) were used to test efficacy of the RNA interference (RNAi) lines. Unless otherwise specified, experimental flies were heterozygous for transgenes. Control heterozygote crosses were performed with consideration to the respective genetic background of the trans-heterozygous Gal4 > UAS flies.
