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. 2019 Mar 12;20(5):1244. doi: 10.3390/ijms20051244

Figure 1.

Figure 1

Doxorubicin induces senescence of breast normal and cancer cells. (A) MDA-MB-231 cells and MCF-10A cells were treated with indicated doses of doxorubicin for 72 h. Cells were either assayed for viability (left panel) or fixed and stained for Ki-67 (right panel, green). Cell viability was determined by WST-1 method and normalized with nontreated con cells. (B–D) Cells were treated with 100 nM doxorubicin for 72 h to induce senescence. Then, cells (B) were incubated with Lysotracker Red (200 nM) for 1 h. Cells (C) were fixed and stained for SA-β-gal. The upper panel shows the bright-field images. The lower panel was the percentage of positive cells (>200 cells scored). Cells (D) were incubated with Mitotracker Red (100 nM) for 30 min. Blue, DAPI stained nuclear. Representative images were captured by a Fluorescence Microscope (100× for A,C; 200× for (B,D). (E) Then days after senescence induction, nontreated con and senescent cells were immunostained with 53BP1, a DNA-SCAR marker. The number of the foci was determined by CellProfiler. Shown was the percentage of cells with >5 foci. (F) Extracts from nontreated con and senescent cells were measured for the indicated proteins by western blotting. Beta-actin was used as the loading control. *** indicates p < 0.001 versus nontreated con.