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. 2021 Jan 6;8(5):2002738. doi: 10.1002/advs.202002738

Figure 2.

Figure 2

VSMC premature senescence and phenotypic switching contribute to plaque vulnerability in CKD/ApoE−/− mice. A) Representative images of HE, α‐SMA, and CD31 staining in aortic root plaques of Sham/ApoE−/− and CKD/ApoE−/− mice. The dashed lines delineate the fibrous cap area. The box indicates the region magnified in the right panels. The arrow indicates fibrous cap VSMCs (α‐SMA+CD31−). The arrowhead indicates endothelial cells (α‐SMA−CD31+). Scale bar (HE), 100 µm. Scale bar (α‐SMA staining), 40 µm. Scale bar (α‐SMA and CD31 double staining), 10 µm. B,C) Quantification of the fold change of fibrous cap thickness and fibrous cap VSMC (α‐SMA+CD31−) contents in aortic root plaques of Sham/ApoE−/− and CKD/ApoE−/− mice (n = 10). D) Representative FC analysis of Ki‐67, SAβG, and α‐SMA expression levels in aortic VSMCs of Sham/ApoE−/− and CKD/ApoE−/− mice. E) The percentages of Ki‐67+ and SAβG+ VSMCs, as well as, the mean fluorescence intensity (MFI) of α‐SMA in aortic VSMCs of Sham/ApoE−/− and CKD/ApoE−/− mice (n = 6). F) Representative images of HE, p53, p16, and γ‐H2AX staining in fibrous cap VSMCs of Sham/ApoE−/− and CKD/ApoE−/− mice. Scale bar (HE), 100 µm. Scale bar (IF), 10 µm. G) Representative transmission electron microscopy images of fibrous cap VSMCs in aortic root of Sham/ApoE−/− and CKD/ApoE−/− mice. The dashed lines delineate the fibrous cap VSMCs. The box indicates the region magnified in the right panels. The arrow indicates the swollen and vacuolated mitochondria in fibrous cap VSMCs. Scale bar (left panels), 10 µm. Scale bar (middle panels), 2 µm. Scale bar (right panels), 1 µm. H,I) The percentages of Ki‐67+ and SAβG+ VSMCs, as well as, the MFIs of p53, p16, and TAGLN in arterial VSMCs of healthy people and CKD patients (n = 10). J) Relative mRNA expression levels of p53, p16, α‐SMA, CNN1, MYH11, and TAGLN in arteries from healthy people and CKD patients (n = 10). Data represent mean ± SD. **p < 0.05, **p < 0.01, two‐tailed Student's t‐test. DAPI, 4,6‐diamidino‐2‐phenylindole; NC, negative control.