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. 2023 Apr 24;8(8):e166643. doi: 10.1172/jci.insight.166643

Figure 7. Knockdown of p16 suppresses RLDC-induced tubular cell senescence and fibrotic phenotype transformation.

Figure 7

BUMPT cells were transfected with p16-shRNA or negative control (NC) shRNA and then selected with puromycin for 2 weeks to establish stable cells for the subsequent RLDC treatment. (A) RT-PCR analysis of p16 mRNA expression (n = 4 experiments). (B and C) Immunoblot and densitometry analysis for P16 protein (n = 3 experiments). (D) Representative images of SA-β-gal staining. Scale bar: 50 μm. (E) Quantification of SA-β-gal–positive cells (n = 3 experiments). (F) Immunoblot analysis for FN, COL1, VIM, and α-SMA. (G–J) Densitometry of FN, COL1, VIM, and α-SMA expression (n = 3 experiments). Quantitative data are presented as mean ± SD. Two-way ANOVA with multiple comparisons was used for statistics. *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001.