Fig. 3

The Ks in the ICD of Dl are required for efficient endocytosis. A–A’’ Clonal analysis of Dl^attP-DlK2R-HA. The homozygous Dl^attP-DlK2R-HA clones are labelled by loss of GFP and highlighted by the arrows. The comparison with the clone homozygous for endogenous Dl (arrowhead), reveals the higher abundance of DlK2R-HA in the apical plasma membrane of homozygous cells (arrows). See also the z-sections in A’, A’’’’, asterisk. A’’ Pixel density measurement of the apical region highlighted in A with the rectangle (s: start-point of measurement). It shows the higher abundance of DlK2R-HA in the apical membrane. B–B’’ Western-blot analysis of the Dl variants revealed that they are similarly expressed (n = 3). C, C’ Detection of surface Dl by applying the primary antibody in the absence of detergence. In this clonal analysis, homozygous Dl^attP-Dl-HA clones (dark green, arrowhead) adjacent to homozygous Dl^attP-DlK2R-HA clones (arrow) were induced and the Dl-variants detected with anti-Dl antibody the binds to the ECD. The staining reveals that homozygous Dl^attP-Dl-HA cells have less Dl on its surface than homozygous or heterozygous Dl^attP-DlK2R-HA cells. D–E’’’’ Wing discs where Dmon1 and Dl^attP-Dl-HA- (D–D’’’’) or Dmon1 and Dl^attP-DlK2R-HA- (E–E’’’’) clones were induced. D, E Overview of the disc bearing the clones. The homozygous Dmon1Dl^attP-variants clones are labelled by the loss of GFP and highlighted by the arrow. D’–D’’’’, E’–E’’’’ z-section of the regions highlighted in D, E by the rectangle. A double mutant clone is outlined in white or yellow. F Quantification of the association of the enlarged Notch-positive endosomes of Dmon1 cells with the HA-signal (Dl-variants, n = 3 for each genotype, see M&M for details). It confirms less association of DlK2R-HA with the enlarged Notch positive endosomes compared to Dl-HA