Research Paper

Figure 1. Extracellular histones promote VSMC senescence and the inflammatory response. VSMCs were treated with various concentrations of histones (0, 10, 25, 50, 100 and 150 μg/mL). (A) The CCK-8 assay was performed to determine cell viability. (B) Cells were treated with 100 μg/mL histones, and the CCK-8 assay was performed to determine cell viability at different time points. (C) SA β-gal staining was used to evaluate cell senescence. (D) Quantitative analysis of SA β-gal-positive VSMCs. (E) Western blotting was performed to analyze p16, p21, and p53 protein expression. (F) Quantitative analysis of (E). (G–I) RT-qPCR was performed to determine the expressions of inflammatory cytokines IL-β, TNF-α, and IL-18. For (A, B, D, and F–I), data are from three independent experiments; mean ± SEM; Student's t-test, ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001 vs. the corresponding control.