Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

Figure 5 MDM2 silencing attenuates anti-senescence effects of formononetin. We silenced the MDM2 gene in MPC-5 cells to investigate whether formononetin (FN) exerts its anti-senescence effects through MDM2 (n = 3 per group). A: Western blot bands of p53 signaling pathway; B: Dual-luciferase reporter gene assay demonstrating that MDM2 silencing attenuates inhibitory effects of FN on p53 transcriptional activity; C-G: Western blot analysis showing that MDM2 silencing abolishes regulatory effects of FN on MDM2, p53, p21, proliferating cell nuclear antigen and CCND1 protein expression; H-J: Silencing MDM2 abolishes inhibitory effects of FN on interleukin-1β, interleukin-6, and tumor necrosis factor α levels in the culture supernatant of high glucose-exposed MPC-5 cells. ^aP < 0.05 vs control groups; ^bP < 0.01 vs control groups; ^cP < 0.05 vs high glucose groups; ^dP < 0.01 vs high glucose groups; ^eP < 0.05 vs short hairpin RNA targeting MDM2 group; ^fP < 0.01 vs short hairpin RNA targeting MDM2 groups. NS: Not significant; FN: Formononetin; HG: High glucose; C: Control; shNC: Short hairpin negative control; shMDM2: Short hairpin RNA targeting MDM2; PCNA: Proliferating cell nuclear antigen; IL-1β: Interleukin-1β; IL-6: Interleukin-6; TNF-α: Tumor necrosis factor α.