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

Figure 8. Both pharmacologic and genetic inhibition of senescence alleviates the paracrine effect of senescent tubular cells on fibroblast proliferation.

Figure 8

(A) Schematic diagram of treatment. Conditioned medium was collected from either post-RLDC BUMPT cells (R-CM) or post-RLDC BUMPT cells treated with ABT-263 (RA-CM). NRK-49F fibroblasts were incubated with serum-free medium containing R-CM or RA-CM for 48 hours for morphological and immunoblot analysis (n = 10 experiments). (B) Cell morphology of NRK-49F. Scale bar: 400 μm. (C) Quantification of the fold changes of NRK-49F fibroblast cell numbers. (D) Immunoblot analysis for FN and VIM in NRK-49F fibroblasts. (E and F) Densitometry of FN and VIM expression. (G–K) Conditioned medium was collected from post-RLDC BUMPT cells stably transfected with p16-shRNA (p16–R-CM) or negative control shRNA (NC–R-CM). NRK-49F fibroblasts were incubated with serum-free medium containing NC–R-CM or p16–R-CM for 48 hours for morphological and immunoblot analysis (n = 4 experiments). (G) Cell morphology of NRK-49F. Scale bar: 400 μm. (H) Quantification of the fold changes of NRK-49F fibroblast cell numbers. (I) Immunoblot analysis for FN and VIM in NRK-49F fibroblasts. (J and K) Densitometry of FN and VIM expression. Quantitative data are presented as mean ± SD. For statistics, 2-tailed, unpaired t test was used. *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001.