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. 2017 Mar 9;15(5):2595–2603. doi: 10.3892/mmr.2017.6303

Figure 2.

Figure 2.

Role of PAR1 in heat stress-induced apoptosis in HUVECs. (A) NC siRNA, PAR1 siRNA, Ad-empty or Ad-PAR1 were transfected into HUVECs, thus achieving knockdown and overexpression of PAR, as confirmed by western blot analysis. (B) Transfected HUVECs were incubated at 37°C (control) or 43°C (heat stress) for 90 min, followed by a 12-h recovery period at 37°C. Apoptosis of HUVECs was analyzed using annexin V and propidium iodide, and the percentage of early apoptotic cells (lower right quadrant) was calculated. (C) Untransfected HUVECs were pretreated with DMSO, 40 µM TF for 10 min or 150 nM SCH for 1 h prior to incubation at 37°C (control) or 43°C (heat stress) for 90 min, followed by a 12-h recovery period at 37°C. Apoptosis of HUVECs was analyzed using annexin V and propidium iodide, and the percentage of early apoptotic cells (lower right quadrant) was calculated. Data are presented as the mean ± standard deviation of three separate experiments. *P<0.05; **P<0.01. PAR1, protease-activated receptor 1; HUVECs, human umbilical vein endothelial cells; NC, negative control; siRNA, small interfering RNA; Ad, adenovirus; DMSO, dimethyl sulfoxide; TF, TFLLR-NH2; SCH, SCH79797; HS, heat stress; Cont, control.