Figure 2. Inhibition of VSMC proliferation by miR-26a.

(A,B) Rat jugular vein VSMCs were transfected with agomir NC, miR-26a agomir, antagomir NC or miR-26a antagomir at different concentrations for 48 h, followed by analysis with quantitative reverse-transcription polymerase chain reaction (qRT-PCR) for miR-26a level. (C,E) Rat jugular vein VSMCs transfected with miR-26a agomir (50 nM) or agomir NC (50 nM) were stimulated with or without PDGF-BB (20 ng/ml) for 24 h. The proliferation of VSMCs transfected with miR-26a agomir was significantly inhibited after stimulation with PDGF-BB in rat jugular veins, as determined by CCK8 proliferation assays and BrdU incorporation. (D,F) Rat jugular vein VSMCs transfected with miR-26a antagomir (100 nM) or antagomir NC (100 nM) were stimulated with or without PDGF-BB (5 ng/ml) for 24 h. The proliferation of rat jugular vein VSMCs transfected with miR-26a antagomir was enhanced by PDGF-BB as determined by CCK8 proliferation assays and BrdU incorporation. (G) Rat jugular vein VSMCs transfected with miR-26a agomir (50 nM) or agomir NC (50 nM) were stimulated with PDGF-BB (20 ng/ml) for 24 h followed by analysis of PCNA and CyclinD1 protein levels with western blotting. The original images for (G) can be seen in Supplementary Fig. 1. (H) Densitometric analysis of PCNA and CyclinD1 protein levels as determined by western blotting. (I) Rat jugular vein VSMCs transfected with miR-26a antagomir (100 nM) or antagomir NC (100 nM) were stimulated with or without PDGF-BB (5 ng/ml) for 24 h followed by analysis of PCNA and CyclinD1 protein levels with western blotting. The original images for (I) can be seen in Supplementary Fig. 4. (J) Densitometric analysis of PCNA and CyclinD1 protein levels as determined by western blotting. *P < 0.05 represents statistical significance compared with the negative control without PDGF-BB, ^#P < 0.05 represents statistical significance compared with the negative control with PDGF-BB (n = 4).