Figure 3.

Effect of has-circ-000595 knockdown on hypoxia-induced damage of V-SMCs. (A) Knockdown efficiency of two siRNAs targeted to has-circ-000595 in V-SMCs. The data are expressed as the mean ± standard deviation. ^*P<0.05 vs. scrabled control. (B) Flow cytometric detection of the apoptotic rate of V-SMCs transfected with siRNAs. The two siRNAs targeted to hsa-circ-000595 did not affect the apoptotic rate of V-SMCs in the absence of CoCl₂; however, under CoCl₂-induced hypoxia, the hsa-circ-000595 siRNAs decreased the apoptotic rate of V-SMCs compared with that of CoCl₂-treated controls. (C) Tunel analysis of the apoptotic rate of V-SMCs transfected with siRNAs in the absence or presence of CoCl₂. The amount of red fluorescence is indicative of the apoptotic rate (magnification, ×20). (D) Expression levels of caspases 3, caspase 8 and Bcl-2 in scramble-, siRNA1- and siRNA2-transfected cells with or without CoCl₂ were detected by western blot analysis, and GAPDH was used as a loading control. siRNA, small interfering RNA; V-SMC, human aortic smooth muscle cell; circRNA, cyclic RNA; hsa, Homo sapiens; Bcl-2, B-cell lymphoma 2; Tunel, terminal deoxynucleotidyl transferase dUTP nick end labeling; PI, propidium iodide; FITC, fluorescein isothiocyanate.