Figure 4: Wurst is involved in clathrin-mediated endocytosis
(A-D) Immunofluorescent analysis using anti-clathrin, anti-Chc, anti-Hsc70,
anti-WurstC (recognizing the C-terminus [13]), and anti-WurstN (recognizing
the N-terminus, see materials and methods) antibodies in Drosophila SL2
cells (A,B) and in membrane sheets of SL2 cells (C,D). The anti-clathrin antibody
recognizes Chc and Clathrin light chain. (A,B) Wurst is localized in a
vesicular like pattern in SL2 cells, which partially co-localize (arrowheads) with
Hsc70 (A) and clathrin (B). (C) Wurst appears distributed in small clusters in
the membrane sheet of a SL2 cell. Wheat germ agglutinin (WGA) selectively
recognizes sugar residues at the plasma membrane surface. White dashes
mark the membrane sheet. (D) Magnification of the membrane sheet reveals
Wurst (green) and Chc (red) overlap (rectangles). (E,F) Confocal images using
anti-Chc and anti-α-Spectrin antibodies in Drosophila stage 17 embryos.
The α-Spectrin marks cell membranes. In wild-type tracheal cells Chc is found
at the membrane and in addition in the cytoplasmic cell cortex (arrowhead in
E). In contrast, in btlG4 driven UAS-RNAi-wurst tracheal specific knock-down
embryos, Chc is accumulated at the apical plasma membrane of tracheal cells
(arrowheads in F). Scale bars indicate 5μm..