Fig. 1

NRP-1 overexpression in BT-474 cell line triggers tumorigenic features. a. Comparison of NRP-1 expression in three breast cancer cell lines MDA-MB-231, MCF-7 and BT-474. NRP-1 was stably overexpressed in the BT-474 cell line and confirmed at the level of protein expression by western blotting and immunofluorescence staining (40× magnification, scale bar 10 μm). b. Wound healing assay images (5× magnification, scale bar 500 μm) taken on day 0 and day 3 indicate increased migration in BT-474 NRP-1 cells compared to control cells 3 days post wound generation. c. NRP-1 overexpression reduced spheroid formation (20× magnification, scale bar 100 μm). d. Western blotting indicated decreased expression of mature form of E-cadherin (lower band) and β-catenin along with e, significantly increased vimentin and f, NRP-1 gene expression. Gene expression is relative to the control BT-474 and normalized to β-Actin and GUSB reference genes expression. The graph represents the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value < 0.05 considered as statistically significant ** p < 0.01