Figure 4. MiR-34b inhibited vascular calcification in mice. (A) Alizarin Red S staining showed calcified arteries in 5/6 NTP+vehicle mice but not SOR mice, and calcification was attenuated in 5/6 NTP+agomiR-34b mice. n = 5. Representative images were shown. The arrows indicate calcification sites in the mouse artery. (B) qRT–PCR showed the expression of miR-34b in SOR group mice, 5/6 NTP+vehicle mice and 5/6 NTP+agomiR-34b mice, respectively. (C) BSP showed the methylation rate of CpG sites of miR-34b DNA in arteries from 5/6 NTP+vehicle mice, was significantly higher than that from SOR arteries. n = 3. Representative images were shown. (D–F) The levels of DNMT3a and Notch1 were determined by western blot and immunohistochemistry staining analysis among arteries from the SOR group, 5/6 NTP+vehicle mice and 5/6 NTP+agomiR-34b mice, respectively. The data were expressed as mean ± SD, *p < 0.05. SOR: sham operation; 5/6 NTP: 5/6 nephrectomy with a high-phosphate diet.