Identification of a new allele of O-fucosyltransferase 1 involved in Drosophila intestinal stem cell regulation

Flies were maintained on standard media at 25°C. Crosses were raised at 18°C in humidity controlled incubators, or as otherwise noted. Flies hatched in 18°C incubators (2–3 days old) were picked and transferred to 29°C incubator, unless otherwise specified. Flies were transferred to new vials with fresh food every day, and dissected at time points specified in the text. In all experiments, only the female posterior midgut was analyzed. Information for alleles and transgenes used in this study can be found either in FlyBase or as noted: stumMB01421 (BL23736), stum204 BL58774), stum4487 (BL58775), stumGal4 (BL58776 and BL58776), Cyo, 2XTb (gift from A. Zhu) (BL36336), UAS-GFP-stum (BL58778) (Desai et al., 2014), pen-2MI02639 (BL36019), UAS-NotchFL (BL26820), UAS-NICD (Cooper and Bray, 2000), Ote+t5.9 (OteP, gift from Chen DH) (Jiang et al., 2008), Df(2R) deficiency kit collections (gift from J. Pastor and from Bloomington), O-fut1SH2260 (BL51666), O-fut1+t3.8 (BL44244), UAS-O-fut1 (BL9376) (Okajima and Irvine, 2002), esgGal4, UAS-GFP, tubGal80ts (esgts, gift from N. Perrimon), DlGal4, UAS-GFP, tubGal80ts (Dlts, gift from S. Hou and R. Xi) (Zeng et al., 2010), GBE+Su(H)Gal4, UAS-GFP, tubGal80ts (GBE+Su(H)ts) (Lu and Li, 2015b), vps26RNAi (THU3819/HMS01769), vps35RNAi (THU3886/HMS01858), NotchRNAi (BL33611), O-fut1RNAi (THU2167/JF02052), FRT19A-Notch264-39, Gbe+Su(H)-lacZ (gift from S. Bray) (Furriols and Bray, 2001), Caf1-p10536 (gift from R. Jiao) (Yu et al., 2013), hsFlp, ActGal4, UAS-GFP; FRT42D-tubGal80 (for MARCM clonal analysis), UAS- GFP.KDEL (BL9898).

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