Research Paper Volume 15, Issue 23 pp 13581—13592

Figure 1. Identification of CaV blockers as a potential new class of senolytics. (A) Schematic representation of the experimental strategy used to identify molecules able to decrease the quantity of CSC-induced senescent cells. (B) Summary of the procedure used to validate which molecules among the 26 identified in (A) display senolytic activity on CSC-induced senescent cells. (C) Dose response curve of CaV blocker Benidipine senolytic activity. X axis shows Benedipine contrations used, y axis shows pourcentage of cell count relative to control DMSO treated cells (% of counted nuclei) (mean ± SEM). (D) After paraformaldehyde fixation, cells were crystal violet stained to visualize cell density. (E) Cells were collected, incubated with trypan blue before being counted. The percentage of dying cells (trypan blue-positive) is shown. (mean ± SEM, One-way ANOVA statistical test, n = 5). (F) Mice were treated once with elastase or vehicle and five time a week with Benidipine or vehicle. Three weeks after elastase injection, lungs were fixed, embedded and slides stained by hematoxylin/eosin. The left panel shows representative images of lungs for each experimental condition. The right panel shows quantification of emphysema by calculating the mean linear intercept. (mean ± SEM, One-way ANOVA statistical test, ctrl: n = 7; elastase: n = 7; Benidipine: n = 3; elastase + Benidipine: n = 5). (G) RT-qPCR results performed on RNA samples extracted from lungs of mice treated or not with Elastase and/or Benedipine. ^*p < 0.05; ^****p < 0.0001.