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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.
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