Fig. 3
NRP-1 promotes the proliferation of pancreatic cancer cells in vitro. a BxPC-3, BxPC-Sc and BxPC-NRPlow cells were cultured for 96 h and cell viability was measured at indicated time points. b, c The above cells harvested after a 48-h culture were subjected to immunoblotting for detecting protein expression as indicated (b), and the density of each band was normalized to β-actin (c). d, e BxPC-3 and BxPC-NRPlow cells harvested 48 h after culturing were subjected to flow cytometry for measuring cell cycle distribution (d) and percentages of cells at different phases were plotted (e). f, g The above cells were stained by EdU (5-ethynyl-2′-deoxyuridine). f Representative images were from EdU-stained nuclei of proliferating cells (green) and DAPI-stained nuclei of all cells (blue), and merged photographs. Magnification bar = 20 µm. g The percentage of EdU-positive cells was plotted. “**” (P < 0.001) indicates a significant difference from BxPC-3 cells