Research Paper
Extracellular histones induce inflammation and senescence of vascular smooth muscle cells by activating the AMPK/FOXO4 signaling pathway
Figure 3. FOXO4 is a downstream target of histone-regulated senescence and inflammation in VSMCs. (A) VSMCs were treated with or without histones (100 μg/mL), and RT-qPCR was performed to determine candidate gene expression. (B–D) VSMCs were treated with various concentrations of histones, and FOXO4 mRNA expression was determined using RT-qPCR (B) or western blotting (C and D). (E) VSMCs were treated with various concentrations of histones, and double immunofluorescence staining was performed to determine FOXO4 and p21 expressions. (F) Quantitative analysis of the fluorescence intensity of FOXO4 and p21 from (E). For (A, B, C, and F), 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.