Extended Data Fig. 4: In vitro cell assays identify PCC1 as a broad-spectrum senolytic.
From: The flavonoid procyanidin C1 has senotherapeutic activity and increases lifespan in mice
(a) Representative images displaying SA-β-Gal staining results after treatment of PSC27 cells with increasing concentrations of PCC1. Scale bar, 30 μm. Data are representative of 3 independent experiments. (b) Senolytic activity appraisal by measuring the percentage of surviving senescent PSC27 cells induced by replicative exhaustion (RS) at increasing PCC1 concentrations. (c) Caspase 3/7 activity-based apoptotic evaluation of CTRL and RS cells treated by PCC1. (d) Senolytic activity assessment by determining the percentage of surviving senescent cells induced by oncogenic HRasG12V (OIS) at increasing PCC1 concentrations. For b and d, P values were calculated by two-sided t-test. (e) Caspase 3/7 activity-based apoptotic appraisal of CTRL and OIS cells treated by PCC1. (f-h) Quantification of the viability of cells in CTRL, TIS, RS and OIS groups treated by 100 μM PCC1 for 3 d. (f), WI38. (g), HUVEC. (h), MSC. The significance of P values for the first concentration of PCC1 that differentiating the survival of CTRL and SEN cells was marked per cell line-based assay. For f, g and h, P values were calculated by one-way ANOVA with Tukey’s multiple-comparison test. (i) Quantification of cell survival in CTRL and SEN populations after combined treatment of 100 μM PCC1 with a ferroptosis inhibitor (liproxstatin-1), necroptosis inhibitor (Nec-1s), caspase-1 inhibitor (VX-765), or a pan-caspase inhibitor (QVD-O-Ph). Cell survival was presented as comparative data of different concentrations of chemicals relative to cells treated by the vehicle. P values were calculated by two-sided t-test. All data are shown as mean ± SD, and are representative of 3 biological replicates. ^, P > 0.05. *, P < 0.05. **, P < 0.01. ***, P < 0.001. ****, P < 0.0001.
