Figure 3

Inhibition of ERα expression by ERα siRNA abolished formononetin-induced stress fiber formation and cell migration in HUVECs.

HUVECs were transfected with non-specific siRNA for 2 days followed by (A) 0.1% DMSO, (B) 12.5, (C) 25 and (D) 50 μM formononetin for 8 h to serve as negative control. HUVECs were transfected with ERα siRNA for 2 days followed by (E) 0.1% DMSO, (F) 12.5, (G) 25 and (H) 50 μM formononetin for 8 h. Cell nuclei were labeled with Hochest 33342 and F-actin was labeled with TRITC-phalloidin. Yellow arrowhead indicated stress fibers terminated at pointed edges. (I) Real-time cell migration of HUVECs transfected with ERα siRNA with or without formononetin treatments as detected by the xCELLigence system. HUVECs were transfected with non-specific siRNA or ERα siRNA for 2 days followed by 0.1% DMSO, 12.5, 25 or 50 μM formononetin for 24 h. (G) Statistical analysis of cell migration of HUVECs transfected with ERα siRNA following formononetin treatment for 20 h. Results are expressed as percentages of controls in mean ± SD (n = 3), **p < 0.01, ***p < 0.001 vs. control.