Fig 1: Effect of miR-142-5p on the mRNA and protein expression levels of MKL2, MMP2 and MMP9. (A) The relative mRNA expression level of MKL2. (B) The relative mRNA expression level of MMP2. (C) The relative expression level of MMP9. (D) Western blot analysis was employed to measure the protein expression levels of MKK2, MMP2 and MMP9. The quantified result of (E) is presented. *P<0.05, inhibitor + siRNA-MKL2 vs. inhibitor group; **P<0.01, inhibitor vs. NC group and inhibitor + siRNA-MKL2 vs. inhibitor group. Control, untransfected cells; NC, miR-142-5p inhibitor negative control-transfected cells; inhibitor, miR-142-5p inhibitor-transfected cells; siRNA-MKL2, cells were transfected with siRNA-MKL2; MKL2, myocardin-like protein 2; MMP2, matrix metalloproteinase-2; MMP9, matrix metalloproteinase-9; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Fig 2: miR-142-5p expression is upregulated, while MKL2 expression is downregulated in the serum of patients with AS. (A) The expression level of miR-142-5p in the serum of 35 patients with AS and 35 healthy volunteers. (B) The mRNA expression level of MKL2 in the serum of 35 patients with AS and 35 healthy volunteers. (C) Results of correlation analysis between the expression of miR-142-5p and the expression of MKL2 in patients with AS. (D) The protein expression level of MKL2 in the patients with AS. (E) The quantified protein expression levels of MKL2 in patients with AS. **P<0.01, AS vs. control group. Control, the serum of 35 healthy volunteers; AS, the serum of 35 patients with AS; AS, atherosclerosis; MKL2, myocardin-like protein 2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Fig 3: MKL2 is identified as a target of miR-142-5p. (A) Target sequences of miR-142-5p in 3'-UTR of the wild-type and mutant MKL2. (B) A dual-luciferase reporter assay was employed to determine whether MKL2 was a target of miR-142-5p. **P<0.01, inhibitor vs. NC group. UTR, untranslated region; WT, wild-type; MKL2, myocardin-like protein 2.
Fig 4: Downregulation of miR-142-5p inhibits PDGF-BB-induced HASMC migration measured by wound scratch assay. (A) Wound scratch assay (magnification, ×200). (B) The quantified result of (A) is presented. **P<0.01, inhibitor vs. NC group and inhibitor + siRNA-MKL2 vs. inhibitor group. Control, untransfected cells; NC, miR-142-5p inhibitor negative control-transfected cells; inhibitor, miR-142-5p inhibitor-transfected cells; siRNA-MKL2, cells were transfected with siRNA-MKL2; PDGF-BB, platelet-derived growth factor type BB; HASMC, human aortic smooth muscle cell; MKL2, myocardin-like protein 2.
Fig 5: Transfection efficiency of miR-142-5p and MKL2. (A) The expression level of miR-142-5p before and after treatment with PDGF-BB. **P<0.01, PDGF-BB vs. before group. (B) The relative expression level of miR-142-5p following transfection with miR-142-5p inhibitor. (C) The relative mRNA expression level of MKL2 following transfection with siRNA against MKL2 (siRNA-MKL2). (D) The relative protein expression of MKL2 following transfection with siRNA-MKL2. (E) The protein/GAPDH expression levels of MKL2 following transfection with siRNA-MKL2 are presented. **P<0.01, siRNA-MKL2 vs. siRNA group. Before, cells before treatment with PDGF-BB; PDGF-BB, cells treated with PDGF-BB; Control, untreated cells; NC, miR-142-5p inhibitor negative control-transfected cells; inhibitor, miR-142-5p inhibitor-transfected cells; siRNA, cells transfected with siRNA; siRNA-MKL2, cells transfected with siRNA-MKL2; PDGF-BB, platelet-derived growth factor type BB; MKL2, myocardin-like protein 2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Supplier Page from Abcam for Anti-Mkl2 antibody