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. 2015 Nov 16;5:16815. doi: 10.1038/srep16815

Figure 4. Blockade of ERα activity by ER antagonist ICI 182,780 abolished formononetin-induced stress fiber formation and cell migration in HUVECs.

Figure 4

HUVECs were treated with (A,a) 0.1% DMSO for 8 h as negative control and (B,b) 50 μM formononetin for 8 h as positive control. For the experiments with ICI 182,780, HUVECs were first treated with ICI 182,780 (10 μM) for 2 h, followed by washout and the treatments of (C,c) 0.1% DMSO, and (D,d) 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. Cortical actin complexes (white arrowhead) were formed in the ICI 182,780 treatment group. (E) Real-time cell migration of HUVECs by using the xCELLigence system showed that ICI 182,780 inhibited formononetin-induced cell migration. HUVECs were treated with ICI 182,780 (10 μM) for 2 h, followed by washout and treatment with 0.1% DMSO or 50 μM formononetin for 24 h. (F) Statistical analysis of HUVECs cell migration following formononetin treatment for 20 h. Results are expressed as percentages of controls (means ± SD; n = 3), **p < 0.01, ***p < 0.001 vs. control.