Endocytic defects in awd mutant cells. Notch trafficking assay was performed on wing discs from (A, C)yw; en2.4-Gal4e22c, UAS-flp/+; Ubi-GFP, FRT82B/FRT82B, awdj2A4 and (B)yw third-instar larvae. (A) In a disc containing awd mutant clone (GFP-negative cells) without synchronized induction of endocytosis (steady state time 0), Notch is seen over-accumulating on or near the cell surface. (B) In wild-type (yw) discs, at 20 minutes after initiation of endocytosis, Notch is expressed at a low level in punctates that are mostly localized in Avl-positive vesicles with some localization in Rab7-positive vesicles (arrows). At one hour after initiation of endocytosis, Notch is barely detectable. (C) In awd mutant clones, at 20 minutes after initiation of endocytosis, Notch over-accumulates exclusively in Avl-positive vesicles but not in Rab7-positive vesicles. This pattern persists at one hour after initiation of endocytosis. The accumulated Notch at this time does not overlap with Rab7-positive vesicles (insets). Bars are 5 μm.
Ignesti et al. BMC Biology 2014 12:12 doi:10.1186/1741-7007-12-12