Figure 3

Identification of miR-378 targets in VSMCs. (A) Genes contain evolutionary conserved predicted binding sites of human miR-378c and involved in atherosclerosis were assessed by the regulation of miR-378c in VSMCs stimulated with PDGF-BB using qRT-PCR. Data were presented as mean ± SD, n = 4. *P < 0.05 as compared with PDGF-treated mimic control group. (B) Blood Samd1 protein levels were measured in 20 ACS patients and 10 healthy subjects. Data were presented as mean ± SD. *P < 0.001 as compared between two groups. (C,D) VSMCs stimulated with PDGF-BB were further transfected with miR-378c mimic. Samd1 mRNA and protein levels were measured by qRT-PCR (C) and western blotting (D), respectively. Data were presented as mean ± SD, n = 4. *P < 0.05 as compared with PDGF-treated mimic control group. β-actin serves as a loading control. (E) VSMCs stimulated with PDGF-BB were further transfected with miR-378c inhibitor. Samd1 mRNA level was measured by qRT-PCR. Data were presented as mean ± SD, n = 4. *P < 0.05 as compared with PDGF-treated vehicle group. (F) Predicted base pairing between miR-378c and 3′UTR of Samd1 by RNA hybrid software. Wild-type or mutated (mut) 3′UTR of Samd1 sequence inserted into dual-luciferase vector were shown. (G) miR-378c mimics were co-transfected with pSI-Samd1-3′UTR or pSI-Samd1-3’UTR-mut into HEK293 cells followed by dual-luciferase analysis. *P < 0.05 compared between the indicated groups, n = 4. (H) Correlation between blood miR-378c level and blood Samd1 protein level. n = 30. Statistical significance was determined by the chi-square test.