Notch localization to the Dx/ESCRT-0-domain in vivo. (A) Localization of endogenous Rab7 (green), Notch (red), and Hrs (blue) in a third-instar wing disc. (A1 and A2) Zoom and merge of Rab7 (green), Notch (red), and Hrs (blue). Zoomed-in images of endosomes (white boxes in A) showing Notch localization on Hrs positive (arrowheads) and negative (arrows) microdomains on the endosomal surface. (B–D) Distribution of endogenous Notch (purple) and Hrs (green) in wing discs. Notch and Hrs are separated in dx¹⁵² mutant (B) but colocalized in Su(dx)^SP mutant disc (C). (D) Mean Pearson’s correlation coefficient values of Notch and Hrs in yw, dx¹⁵², and Su(dx)^SP. Error bars are SEM, ***P < 0.001 by two tailed t test, sample sizes are indicated in the figure.