Fig. 5.

DCA-induced p38 MAPK activation promotes VSMC calcification via Runx2. A) Runx2 deletion blocked DCA-induced VSMC calcification. VSMC from control and SMC-specific Runx2 deletion mice were exposed to 0 and 5 mM DCA in osteogenic media for 21 days. a) Alizarin red staining for calcification. Representative images of 3 experiments performed in duplicate are shown. b) Quantitative measurement of calcium content in separate sets of experiments (n = 3, *p < 0.001). c) Western blotting analysis of Runx2 expression and p38 MAPK phosphorylation/activation. B) Knockdown of p38 MAPK inhibited Runx2-induced VSMC calcification. Stably selected VSMC with control (sh Scr) or p38 MAPK shRNA (sh p38) were infected with lentivirus carrying control or Runx2 protein, and cultured in osteogenic media for 21 days. a) Alizarin red staining for calcification. Representative images of 3 independent experiments performed in duplicate are shown. b) Quantitative measurement of calcium content in separate sets of experiments. (n = 3, *p < 0.001). c) Western blotting analysis of Runx2 expression and p38 MAPK phosphorylation. Representative results of three independent experiments are shown.