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. 2021 Jun 5;10(6):916. doi: 10.3390/antiox10060916

Figure 8.

Figure 8

Procyanidin B2 attenuates the activation of ERK1/2 on HASMC calcification. (A) HASMCs were transfected with pEGFP-C2 or C2-MEK1-mut for 24 h. The transfected cells had calcification induced in the presence of PB2 (2 μM) for 7 days. The expression of p-ERK1/2, and RUNX2 protein was determined by Western blot. ** p < 0.01, *** p < 0.001 (n = 3). HASMCs were transfected with pEGFP-C2 or C2-MEK1-mut for 24 h and then cultured in a CM medium with PB2 (2 μM) for 2 weeks, followed by alizarin red S staining (B) and the quantitative analysis of stained dye (C). * p < 0.05, *** p < 0.001 (n = 3). (D) HASMC calcification was induced by CM in the presence of PB2 (2 μM) for 2 days, and then the cells were treated with EGF (10 ng/mL) for another 5 days. The expression of p-ERK1/2 and RUNX2 protein was determined by Western blot. ** p < 0.01, *** p < 0.001 (n = 3). HASMCs were cultured in a CM medium containing PB2 (2 μM) for 1 week, and then they received EGF (10 ng/mL) treatment for another week, followed by alizarin red S staining (E) and the quantitative analysis of the stained dye (F). * p < 0.05 (n = 3), ** p < 0.01.