Fig. 2. Monotropein effect on vascular smooth muscle cell proliferation.
(A) CCK-8 assay was conducted to assess the effect of different concentrations of monotropein on vascular smooth muscle cell viability. n = 3 independent experiments. (B) EDU assay to determine the proliferation of vascular smooth muscle cells; scale bar, 100 µm. n = 3 independent experiments. (C) Protein levels of PCNA, α-SMA in cells. n = 3 independent experiments. (D) Calculation of proliferation rate of vascular smooth muscle cells. (E) Quantitative analysis of EDU in vascular smooth muscle cells. (F, G) Quantitative analysis of PCNA and α-SMA. (H, I) Quantification of PCNA and Acta2 expressions by qPCR. (J, K) Immunofluorescence staining of mouse aortic root and quantitative analysis of arterial plaques with PCNA (green) and Myh11 (red). Scale bar, 50 µm. n = 3 independent experiments. (L) Expressions of PCNA and Acta2 in the aorta. n = 3 independent experiments. (M, N) EDU proliferation staining and quantification of mouse aorta; scale bar, 50 µm. n = 3 independent experiments. (O, P) Quantitative analysis of PCNA and Acta2 expressions in the aorta. Bar graphs depict mean ± SEM; one-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. PCNA, proliferative cell nuclear antigen; α-SMA, α-smooth muscle actin; ox-LDL, oxidized low density lipoprotein; HFD, high-fat diet; MTP, monotropein; NCD, normal diet.
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