Research Paper
Extracellular histones induce inflammation and senescence of vascular smooth muscle cells by activating the AMPK/FOXO4 signaling pathway
Figure 4. FOXO4 is involved in extracellular histone-facilitated VSMC inflammation and senescence. (A–C) VSMCs were transfected with shFOXO4-1#, shFOXO4-2#, or shCon vectors; then, RT-qPCR and western blot were performed to determine FOXO4 expression. (D) VSMCs were treated with histones after being transfected with shFOXO4 or shCon vector, and SA β-gal staining was performed to evaluate cell senescence. (E) Quantitative analysis of relative SA β-gal-positive cell numbers from (D). (F and G) RT-qPCR was performed to determine the expressions of the inflammatory cytokines IL-β and TNF-α in VSMCs after the indicated treatment. For (A, C, E, F, and G), data are from three independent experiments; mean ± SEM; Student's t-test, *P < 0.05 and **P < 0.01 vs. the corresponding control.