Fig. 2. Notch was endocytosed but not transported to early endosomes in O-fut1^- cells. (A-K) Live wing discs containing Notch^- (A,B) or O-fut1^- (C-K) clones were incubated with an anti-Notch extracellular antibody (rat1; magenta) to detect Notch on the plasma membrane, and allowed to endocytose Notch for 20 minutes (A-J) or 10 hours (K). Mutant cells were distinguished by the lack of GFP (green, A-F,K) or are labeled O-fut1^- (G-J). Wing discs were also stained with an anti-Hrs antibody (G-J, shown in green). Optical sections corresponding to the apical region (A,C,G) or to 2 µm (D,H) or 8 µm (E,I) beneath the apical level, and optical vertical sections (B,F,J) are shown. (L) Living wing discs with O-fut1^- clones (indicated by O-fut1^-) incubated with fluorescent dextran (green) for 10 minutes, chased for 20 minutes at 25°C, and stained with an anti-Notch antibody (C17.9C6; magenta). Higher magnification of one or two dextran-positive vesicles in wild-type (arrowhead) and O-fut1^- (open arrowhead) cells are shown as insets in the upper left and lower right, respectively. The clone boundaries are demarcated by a white dashed line. All wing discs were isolated from late third-instar larvae.