Research Paper Volume 16, Issue 16 pp 11796—11808

Figure 3. The senescence-modifying effect of 2-BP is detected only if cells are treated before senescence induction. VSMCs were treated with 2-BP for 24 h before (pretreatment) or after (posttreatment) doxorubicin (DOX)-mediated DNA damage induction. DNA damage was analyzed at different time points (3 and 7 days) following incubation with 2-BP. (A) Schematic representation of the experimental design. The graph shows the number of DNA damage-associated nuclear 53BP1 and γH2AX foci after doxorubicin treatment (B, C), the percentage of cells that incorporated BrdU (D), and SA-β-gal activity (E). The results are presented as the means ± SEMs from two separate experiments, each with N=3 biological replicates. Statistical analysis was performed using one-way ANOVA; *p< 0.05, **p<0.01. Representative images of SA-β-gal activity and 53BP1 and γH2AX levels are shown in Panel (F).