Figure 4

hsa_circ_0081065 act as miR-665 sponge to up-regulate HIF-1α in HUVECs. (A) Starbase 2.0 was used to predict the miRNA targets of hsa_circ_0081065; (B) RNA-pull down assay was used to detect the relative expression of hsa_circ_0081065 pulled down by using miR-665 probe; (C) Dual-luciferase reporter assay was used to detect the luciferase activity of wild-type or mutant reporter of hsa_circ_0081065 with miR-665 overexpression; (D) TargetScan was used to detect the binding site of miR-665 to HIF-1α; (E) RNA-pull down assay was used to detect the relative expression of HIF-1α w pulled down w by using miR-665 probe; (F) Dual-luciferase reporter assay was used to detect the luciferase activity of wild-type or mutant reporter of HIF-1α with miR-665 overexpression; (G) The luciferase activity of wild-type reporter of HIF-1α could be reversed by overexpressing hsa_circ_0081065; (H, I) RT-qPCR and Western blot was used to detect the expression of HIF-1α with miR-665 mimics transfection in HUVECs; (J, K) RT-qPCR and western blot was used to detect the expression level of HIF-1α rescued by overexpressing hsa_circ_0081065; ***P < 0.001; ****P < 0.0001.