Figure 5From: Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cellsThe effects of Y27632 on the internalization of the EGFR in Panc1 pancreatic cancer cells. (A) Panc1 cells were treated with the indicated concentrations of Y27632 or vehicle for 1 h at 37°C, and then labeled for 15 min at 37°C with anti-EGFR antibodies which recognize the extracellular domain of the EGFR. The cells were then exposed to 30 ng/ml of EGF for 10 min at 37°C. To observe only the cell surface EGFR that remained on the plasma membrane, these cells were not permeabilized with Triton X-100. They were then exposed to Alexa Fluor 488®-conjugated donkey anti-goat IgG antibodies and DAPI for 1 h. Representative cells examined by fluorescence microscopy are shown. (B) Green fluorescence signals from the remaining cell surface EGFR were measured using the NIH image software program, and expressed as the % of the control (compared with untreated control cells).Back to article page