Figure 1. MiR-34b was down-regulated and involved in the process of VSMCs calcification. (A–C) VSMCs cultured in 3.5 mM of Pi were measured for ALP activity, OC secretion and Runx2 expression. (D) VSMCs were treated with 3.5 mM of Pi for 18 days and then subjected to Alizarin Red S staining. The calcium content was extracted with cetyl-pyridinium chloride and quantified by spectrophotometry. Representative images were shown. The arrows indicate the mineralised nodules. (E) Expression of miR-34b was detected by qRT–PCR in cultured VSMCs. (F) qRT–PCR showed the expression of miR-34b in VSMCs transfected with scramble miRNA mimics, inhibitor or miR-34b mimics, miR-34b inhibitor, respectively. (G–I) VSMCs were transfected to detect the change of ALP activity, OC secretion and Runx2 protein levels. n=3. The data were expressed as mean ± SD, *p < 0.05; **p < 0.005; ***p < 0.0005.