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. 2017 Mar 14;8:111. doi: 10.3389/fphar.2017.00111

Figure 5.

Figure 5

ALK1 receptor gene silencing by siRNA abrogates Smad1/5/8 phosphorylation, VEGF upregulation and angiogenesis induced by Dkk-3. (A) RTPCR analysis of ALK1 mRNA level in both control and ALK1 silenced cells. **p < 0.001 vs. control-siRNA treated cells. (B) Western blot analysis of phospho-Smad1,5,8 expression in both control and ALK1 silenced cells exposed to hrDkk-3 (10 ng/ml, 60 min). HUVECs incubated with TGF-β1 were used as positive controls. Values are means ± SEM. **p < 0.001 vs. baseline conditions, ##p < 0.001 vs. hrDkk-3. Immunocytochemical (C) and western blot analysis (D) of VEGF expression in both control and ALK1 silenced cells exposed to hrDkk-3 (10 ng/ml, 12 h). Values are means ± SEM. **p < 0.001 vs. baseline conditions, #p < 0.05 vs. hrDkk-3, ##p < 0.001 vs. hrDkk-3. (E) Endothelial cell tube formation of both control and ALK1 silenced cells treated with hrDkk-3 (10 ng/ml, 18 h). Values are means ± SEM. *p < 0.05 vs. baseline conditions, #p < 0.05 vs. hrDkk-3.