Fig. 4

Molecular mechanism of CML-accelerated VSMC calcification under high-lipid, apoptosis-coexisting conditions. After the cells reached 80% confluence, A7r5 aortic smooth muscle cells were cultured in the presence of 50 μg/mL oxLDL and 80 μg/mL RAW264.7-derived-ABs with the indicated treatment for 7 days, and the medium was replaced every 2 days. A Presents the expression of related protein in treated A7r5 cells by Western blot. B–D present quantitative analysis of relative RAGE, Nox4 and P-p38 protein levels (target protein/β-actin) by densitometry. Values are expressed as mean ± SD from three independent experiments. a Control group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL); b CML group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL, and 10 μmol/L CML); c anti-RAGE group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL, 10 μmol/L CML, and 100 μg/mL antibody against RAGE); d NADPH oxidase inhibitor group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL, 10 μmol/L CML, and 10 μmol/L DPI); e p38MAPK inhibitor group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL, 10 μmol/L CML, and SB203580); f anti-cbfα1 group (A7r5 medium supplemented with 80 μg/mL RAW264.7-derived-ABs plus 50 μg/mL oxLDL, 10 μmol/L CML, and 100 μg/mL antibody against cbfα1). Values are expressed as mean ± SD from three independent experiments