Research Paper Volume 16, Issue 16 pp 11796—11808

Figure 2. 2-BP treatment decreases the expression of senescence markers in doxorubicin-treated VSMCs. Cells were pretreated with increasing doses of 2-BP for 24 h (pretreatment), washed and induced to senescence by a 2 h treatment with doxorubicin (DOX). The proliferation rate during a 4-day culture measured every 2 h was used to calculate the growth rate (A, B). Changes in the number of BrdU-positive cells (C), the level of SA-β-gal activity (D), and the number of DNA damage-associated 53BP1 (E) and γH2AX foci (F) 3 days after doxorubicin treatment in response to increasing doses of 2-BP. Green line marks the mean level of analyzed parameter in control, untreated cells. The results are shown as the means with SEM from N=3 or 5 biological replicates. Statistical analysis was performed using one-way ANOVA (comparison between DOX and 2-BP + DOX) or two-way ANOVA (for B), p≤ 0.05, **p≤0.01. Representative images of SA-β-gal activity and 53BP1 and γH2AX levels are shown in Panel (G).