FIG. 5.

Effects of Anx2 depletion on the basal rate of EGFR downregulation. (A) Stably transduced COS-1 cells (Scr and Anx2) were seeded in six-well plates and serum starved overnight. The next day, cells were treated with medium containing EGF and cylcoheximide or cycloheximide alone for 10 min. Cells were harvested immediately (0 h) and at 1 and 4 h following treatment. Cell lysates were analyzed by SDS-PAGE and Western blotting with anti-EGFR, anti-Anx2, and anti-actin antibodies. (B) Graphical representation of the experiments. EGFR levels in cells at 4 h following treatment were quantified using Image J software. EGFR levels in cells that received no EGF were normalized to 100. The graph represents two independent experiments.