Figure 5

Effects of D609 on trafficking and degradation of HER2 receptor molecules in SKBr3 cells. (a) CLSM analyses of SKBr3 cells, incubated for the indicated time intervals with the PC-PLC inhibitor D609 (50 μg/mL) and then fixed and stained with the α-HER2 mAb W6/100 (red) and α-Rab5B Ab (green, panels a through d) or α-Lamp 2 Ab (green, panels e through h). Nuclei were stained with DAPI (blue). Scale bars, 8 μm. (b) Western blot analyses of total lysates of SKBr3 cells either untreated or incubated with D609 for 6 and 24 hours. The blots were incubated with α-HER2 or α-PC-PLC Abs. Actin was used as quantitative loading control. Experiments shown in panels a and b were independently repeated 3 and 4 times, respectively.