Effects of ANXA1 knockdown on migration and invasiveness rate of MIA PaCa-2 and PANC-1 cells. Western blot using an anti-ANXA1 antibody on protein extracts from MIA PaCa-2 (panel A) or PANC-1 cells (panel D) treated or not with siRNAs direct against ANXA1 (siANXA1). Δ represents ANXA1 fold change normalized to control levels by densitometry. Protein normalization was performed on tubulin levels. B, Results of wound-healing assay on MIA PaCa-2 (panel B) or PANC-1 cells (panel E) transfected with siANXA1s or scrambled siRNAs. Statistical significance was calculated using unpaired t-test between control and ANXA1 knock-down cells, **p < 0.01 vs untreated control. The data are representative of 5 independent experiments ± SEM. Invasiveness rate of MIA PaCa-2 (panel C) or PANC-1 cells (panel F). In invasion assays a total of 90,000 cells were transfected or not with siANXA1s (5nM) or scrambled siRNAs (5nM) for 72 h and plated as described in Methods section. Invasiveness rate was founded out by counting stained cells on the lower surface of the filters. Data represent mean cell counts of 12 separate fields per well ± SEM of 5 experiments. Statistical significance was calculated using unpaired t-test between control and ANXA1 knock-down cells **p < 0.01 vs serum free (SF) control; ***p < 0.001 vs SF control; ###p < 0.001 vs control.