Fig. 9. Suppression of Beclin expression does not disrupt EGF-stimulated endocytosis or post-endocytic degradation of the EGFR. (A) Control or Beclin KD cells were fixed and processed for immunofluorescence detection of EGFR after overnight growth in serum-free medium (No EGF), and after 30 minutes or 70 minutes incubation with 200 ng/ml EGF. Bar, 10 µm. (B) Cells were harvested at the indicated times after addition of EGF and subjected to immunoblot analysis for total EGFR (upper panel). The bar graph (lower panel) shows the data derived from Kodak 440CF Imager scans performed on blots from three cultures harvested at each time point. (C) Cells were harvested at the indicated times after addition of EGF and subjected to immunoblot analysis for total EGFR (upper panel) or EGFR phosphorylated on Tyr1068 (pEGFR). The bar graph depicts the ratio of pEGFR to total EGFR determined in three parallel control or Beclin KD cultures at 30 minutes after EGF stimulation (mean ± s.e.).