Fig. 3

The constitutive internalization of the exon 19 deletion mutant under steady-state condition is independent of dynamin activity. a, representative confocal sections are shown with magnified insets to illustrate the colocalizations of EGFR with EEA1 and LAMP2 in HCC827 cells. Images were captured using a Leica TCS SP5II microscope with a 63X objective. b, serum-starved A549 cells were treated with EGF at indicated concentrations for 6 h with or without dynasore addition to block the dynamin activity. Cell lysates were analyzed by immunoblottings with indicated antibodies. The column chart below shows the quantification data of EGFR. c and d, HCC827 and H1650 cells were treated with the dynamin inhibitors, dynasore (80 μM) and dyngo-4a (20 μM), for indicated times. Cell lysates were subjected to immunoblotting analysis to detect the levels of EGFR. The column charts below show the quantification data of EGFR expression. e, immunofluorescence analysis of EGFR distribution in HCC827 and H1650 cells treated with dynasore, dyngo-4a, or control reagents. Images were captured using a fluorescent microscope (Olympus BX63, 40X objective). Scale bar = 10 μm. All error bars represent the standard error of the mean (n = 3), and * indicates p < 0.05