Fig. 5

Effects of silencing NE on pVICs calcification. pVICs were transfected with 5 nM siRNA targeted NE or negative control for 24 h. A RT-qPCR detection of siRNA targeted NE knockdown efficiency. B, C RT-qPCR detection of mRNA expression of RUNX2 and OPN in each groups. D–I Western blot analysis of TNF-α, osteogenic markers (RUNX2, OPN) and apoptosis proteins (bcl-2, bax) in each groups. J Calcium deposition was measured by Alizarin red staining in pVICs cultured in osteogenic medium for 14 days. K Early osteogenic differentiation was measured by ALP staining in pVICs cultured in osteogenic medium for 7 days. *P < 0.05, **P < 0.01 vs corresponding control groups. NS meant no significant difference. The measurement data are expressed as the mean ± SD. All experiments were independently repeated three times