Expression and localization of ANXA1 in human PC cells. A, Total ANXA1 expression in MIA PaCa-2, PANC-1, BxPC-3 and CAPAN-2 cells was analyzed by Western blot with anti-ANXA1 antibody. Δ represents ANXA1 fold change by densitometry. B, Cultured human MIA PaCa-2, PANC-1, BxPC-3 and CAPAN-2 cells fixed and labelled with fluorescent antibody against E-cadherin and vimentin (green). C, D, Quantitative analysis of E-cadherin and vimentin protein expression in MIA PaCa-2, PANC-1, BxPC-3 and CAPAN-2 cells. Histograms represent the percentage of E-cadherin and vimentin positive cells on total DAPI positive nuclei. Results are mean ± standard deviation from three independent experiments, where at least 100 nuclei per cell line were counted. E, Immunofluorescence analysis to detect FAK (panels a, d), ANXA1 (panels b, e, h, m), and F-actin (panels g, l) in MIA PaCa-2 and PANC-1. Nuclei were stained with DAPI. The merged image shows overlapping localization of the proteins (panels c, f, I, n). Magnification 63×. The data are representative of 5 experiments with similar results. Bar = 10 μm.