Figure 2

PHB is indispensable for EGF-induced ERK activation in pancreatic cancer cells. A. Immunoblot analysis of PHB and p-ERK1/2 levels in AsPC-1 and Capan-2 cells. α-tubulin was used as a loading control. B. AsPC-1 and Panc-1 cells were transfected with PHB-specific (siPHB) or control (siCon) siRNAs. At 48 h post-transfection, cells were treated with EGF (50 ng/ml) for 15 min and then subjected to immunoblot analysis using the indicated antibodies. p-ERK1/2 was detected as an activation marker and α-tubulin as the loading control. C. Confocal microscopic images of the localization of PHB (red) and p-ERK1/2 (green). AsPC-1 cells were transfected with PHB-specific (siPHB) or control (siCon) siRNAs. After 48 h of transfection, cells were stimulated with EGF (50 ng/ml) for 15 min. AsPC-1 cells were incubated with antibodies specific for PHB and p-ERK1/2. Nuclei were counterstained with DAPI. Scale bars, 10 μm. Results are representative of three independent experiments.