Research Paper Volume 16, Issue 3 pp 2438—2456

Figure 3. Mark4 knockdown inhibited β-GP-induced VSMCs apoptosis and calcification. VSMCs were infected with lentiviral shMark4 (sh-Mark4) or sh-negative control (sh-NC) under high phosphate conditions for 72 h. (A) The RT–qPCR showed transfection efficiency of sh-Mark4a-c in β-GP mediated VSMCs. ^*Means p < 0.05 vs β-GP+ sh-NC group. (B) The inverted fluorescence microscopy observation of sh-Mark4b transfection efficiency in β-GP-treated VSMCs. (C, D) Western blot and quantitative densitometry analysis of Mark4, Akt and apoptosis-related proteins in VSMCs with knockdown of Mark4. (E) Apoptosis of sh-Mark4 transfected VSMCs cultured with β-GP medium was assessed by TUNEL staining (green). Nuclei were counterstained with DAPI (blue). (F) Western blot and quantitative densitometry analysis of Runx2 in VSMCs with knockdown of Mark4. (G) Alizarin Red S staining and (H) calcium content in VSMCs infected with sh-Mark4. ^*Means p < 0.05, ^**p < 0.01, ^***p < 0.001 vs. control, ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 vs. β-GP+ sh-NC. Error bar represents three independent experiments.