Fly Gut Stem Cell Regulation

Fly stocks were maintained at 25°C on standard food under a ∼12 h/12 h light/dark cycle. Food consisted of 15.8g yeast, 9g soy flour, 5.2g agar, 67g cornmeal, and 0.5% propionic acid. To avoid larval overpopulation, <30 adult flies per vial were transferred to new food vials every 2–3 days.
The following stocks were used in this study: esg-Gal4,tub-Gal80ts,UAS-GFP/CyO (esg^ts>GFP (gift from Bruce Edgar) [48 (link)]; FRT82B, tub-Gal80/TM6B (gift from Bruce Edgar) [49 (link)]; Myo1A-Gal4,tub-Gal80ts,UAS-GFP/CyO (Myo1A^ts> GFP) (gift from Bruce Edgar) [48 (link)]; UAS-CncC (gift from Dirk Bohmann) [50 (link)]; UAS-Png1WT; UAS-Png1C303A; Png1ex18; Png1ex14 (gift from Hamed Jafar-Nejad) [28 (link)]; UAS-OGARNAi (VDRC, #106670); UAS-OGARNAi (VDRC, #41822); UAS-OGTRNAi (VDRC, #18610); UAS-OGTRNAi (VDRC, #18611); UAS-Png1RNAi (VDRC, #103607); w¹¹¹⁸ (BDSC, #3605); UAS-Png1RNAi (BDSC, #54853); UAS-ENGase RNAi (BDSC, #64609).
The UAS-OGT line that contains Myc epitope tag in the N terminus (UAS-Myc-OGT) was made by P-element-mediated transformation and OGAdel.1 mutant was generated by standard P-element excision [51 (link)].
For transgene expression at specific developmental stages, the Gal80ts technique was used. The flies were set up and maintained at 22°C until adulthood. After maintaining the flies at 29°C, the midguts were dissected and analyzed.