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. 2023 May 15;22(7):e13865. doi: 10.1111/acel.13865

FIGURE 2.

FIGURE 2

PINK1 deficiency increases senescence and senescence‐associated secretory phenotypes, aggravating renal aging and renal tubular epithelial cells. (a) The mRNA levels of senescence signaling mediators (p53, p16, and p21) in Pink1+/+ and Pink1−/− mice at the age of 4 and 24 months. (b) Western blotting of senescence signaling mediators (p53, p16, and p21) and cell cycle markers (proliferating cell nuclear antigen (PCNA) and p‐Rb) in Pink1+/+ and Pink1−/− mice at the age of 4 and 24 months. (c) The mRNA levels of SASPs (CTGF, Fn, a‐SMA, TGF‐β1, NF‐kb, and IL‐1β) in Pink1+/+ and Pink1−/− mice at the age of 4 and 24 months. (d) Western blotting of SASPs (CTGF, Fn, a‐SMA, TGF‐β1, IL‐1β, NF‐kb, and p‐NF‐kb p65) in Pink1+/+ and Pink1−/− mice at the age of 4 and 24 months. (e) Quantitative protein level of PCNA and p‐NF‐kb/NF‐kb for Western blot assay in Pink1+/+ and Pink1−/− mice kidneys at the age of 4 and 24 months. (f) The mRNA levels of senescence signaling mediators, p53, p16, and p21, in siCONT and siPINK1 renal tubular epithelial cells (HKC‐8) with H2O2 treatment. (g) Western blotting of senescence signaling mediators (p53, p16, and p21) and a cell cycle marker (p‐Rb) in siCONT and siPINK1 H2O2‐treated HKC‐8 cells. (h) The mRNA level of SASPs (CTGF, Fibronectin, α‐SMA, TGF‐β1, and IL‐1β) in siCONT and siPINK1 H2O2‐treated HKC‐8 cells. (i) Western blotting of SASPs (Fibronectin, α‐SMA, TGF‐β1, p‐NF‐kb‐p65, and NF‐kb p65) in siCONT and siPINK1 H2O2‐treated HKC‐8 cells. (j) Quantitative protein level of p‐NF‐kb/NF‐kb for Western blot assay in siCONT and siPINK1 H2O2‐treated HKC‐8 cells. mean ± standard error of mean. (a, c, e) *p < 0.05, **p < 0.01, ***p < 0.001 vs. Pink1+/+ 4 M, # p < 0.05, ## p < 0.01, ### p < 0.001 vs. Pink1−/− 4 M, † p < 0.05, †† p < 0.01, ††† p < 0.001 vs. Pink1+/+ 24 M, M, months. (f, h, j) *p < 0.05, **p < 0.01, ***p < 0.001 vs. siCONT without H2O2, # p < 0.05, ## p < 0.01, ### p < 0.001 vs. siPINK1 without H2O2, † p < 0.05, †† p < 0.01, ††† p < 0.001 vs. siCONT with H2O2, CONT, control.