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. 2023 Jul 12;136(20):2484–2495. doi: 10.1097/CM9.0000000000002763

Figure 1.

Figure 1

Expression of Lin28a and the let-7 family in VSMCs in RS. (A) Expression and colocalization of Lin28a (red) and α-SMA (green) in human post-PTA RS and AS samples. α-SMA is the specific marker of VSMCs. The immunofluorescence image was obtained at magnification ×40 and the scale bar represents 500 µm; a magnification ×400 image is shown in the inset on the bottom right corner of the merged image; the scale bar represents 50 µm. (B) Quantification of Lin28a+ α-SMA+ cells in the neointima area. (C) The expression levels of let-7 miRNAs in human AS and RS (n = 5). P <0.05 vs. AS. (D) The immunohistochemical staining showed the expression of α-SMA in AS and RS plaques in rat models. The image was obtained at magnification ×100 and the scale bar represents 200 µm. (E) Quantification of positive staining for α-SMA in the AS and RS groups. (F) Expression of let-7 miRNAs in rats with AS and PTA-related RS. P <0.05 vs. AS. (G) Western blotting was performed to examine Lin28a expression after transfection with up- or down-regulating Lenti-virus in cultured primary VSMCs. (H) Statistical analysis of Western blotting results displayed by the bar graph. P <0.05 vs. Lenti-NC-shRNA group. qRT-PCR was performed to examine let-7 miRNAs after transfection with up-regulation (I) or down-regulating (J) Lin28a Lenti-virus in VSMCs. The results showed that let-7c, let-7g, and miR98 were negatively regulated by Lin28a. P <0.05 vs. Lenti-NC-shRNA group. α-SMA: α-smooth muscle actin; AS: Atherosclerosis; DAPI: 4′,6-diamidino-2-phenylindole; Lenti-NC-shRNA: Normal control short-hairpin RNA lentivirus; miRNA: microRNA; miR98: microRNA98; PTA: Percutaneous transluminal angioplasty; qRT-PCR: Quantitative real-time polymerase chain reaction; RS: Restenosis; shRNA: Short-hairpin RNA; VSMCs: Vascular smooth muscle cells.