CDKN2A/p16INK4a Antibody - #AF5484

Fig. 6: Knockout of cdkn2a (p16) in Lyz2+ cells abrogates LSI-induced endplate sclerosis. a Representative immunofluorescent images of p16+ (red), F4/80+ (green) cells and DAPI (blue) staining of nuclei in mouse caudal endplates of L4/5 in Cdkn2aΔLyz2 or Cdkn2af/f mice at 8 weeks after LSI or sham surgery. Scale bars, 50 μm. b Representative immunofluorescent images of CD31 (green), Emcn (red) and DAPI (blue) staining of nuclei in the endplates of Cdkn2aΔLyz2 or Cdkn2af/f mice after LSI or sham surgery. Scale bars, 50 μm. c Permillage of CD31+Emcn+ area in the endplates of b. d Representative µCT images of the caudal endplates of L4/5 (coronal view) in Cdkn2aΔLyz2 or Cdkn2af/f mice at 8 weeks after LSI or sham surgery. Scale bars, 500 μm. e Quantitative analysis of the total porosity of d. f Representative images of safranin O and fast green staining of endplates in Cdkn2aΔLyz2 or Cdkn2af/f mice at 8 weeks after LSI or sham surgery. Scale bars, 50 μm. g Endplate scores of the endplates of f. h Representative immunohistochemical images of Osterix (Osx) in the endplates of Cdkn2aΔLyz2 or Cdkn2af/f mice at 8 weeks after LSI or sham surgery. Scale bars, 50 μm. i Quantitative analysis of the number of Osx+ cells in the endplates of h. j Representative images of immunofluorescent analysis of CGRP+ sensory nerves (red) and DAPI (blue) staining of nuclei in the endplates of Cdkn2aΔLyz2 or Cdkn2af/f mice after LSI or sham surgery. Scale bars, 50 μm. k Permillage of CGRP+ area in the endplates of l. n = 6 per group. Data are represented as means ± standard deviations, as determined by One-way ANOVA. Source data are provided as a Source Data file.

Figure 1 Cellular senescence is associated with aortic valve calcification. A) GSEA plot showing differential expression of signature genes in the cellular senescence pathway in calcific valve and normal valvular tissue based on the RNA-seq data sets from the GEO database. B) DEG plot showing the differential expression genes between the calcific valve and normal valvular tissue. C) Violin plots representing the expression of P21, P16, and P53 in the aortic valve from CAVD patients, elder and young individuals. D) Representative Vonkossa staining, Alizarin red staining, and immunohistochemical staining of P16, P21, and P53 in aortic valves from CAVD patients and controls. Scale bar 200 µm. E) Immunofluorescent staining of NQO-1 (red), CCND1 (red), and DAPI (blue) in the aortic valve from CAVD patients and controls. Scale bar 100 µm. F) Protein expression of ALP, Runx2, NQO1, and P21 in the aortic valve from CAVD patients (n = 10) and controls (n = 10). Bar plots representing the fold change of specific protein expression over control. Data are means ± SD. **p < 0.01; ***p < 0.001 (unpaired two-tailed Student's t-test).

Fig. 1. Effects of AGEs in different concentrations on the senescence of BMSCs. The BMSCs were treated with AGEs (50–200 μg/mL) or BSA for 24–72 h. (A) SA-β-gal assay for detection of BMSCs senescence. Scale bar: 100 μm. (B) Detection of H3K9me3 by immunofluorescence in BMSCs. Scale bar: 100 μm. (C) Detection of γ-H2AX by immunofluorescence in BMSCs. Scale bar: 100 μm. (D–I) Representative Western blotting assay and quantitation of the level of P16, P21, P53. **p < 0.01 versus BSA.