Fig. 3: Cisplatin-induced DNA damage is not removed along of time in three-dimensional (3D)-cultured MCF-7 cells and correlates with senescence induction. | Cell Death & Disease

Fig. 3: Cisplatin-induced DNA damage is not removed along of time in three-dimensional (3D)-cultured MCF-7 cells and correlates with senescence induction.

From: ATR mediates cisplatin resistance in 3D-cultured breast cancer cells via translesion DNA synthesis modulation

Fig. 3

a–c γ-H2AX immunodetection followed by flow cytometry and d, e cisplatin–DNA adduct detection by slot-blot assays, in cells exposure to 60 μM of cisplatin by different periods of time, for evaluation of DNA damage processing. f, g β-Galactosidase staining at pH 6 in 3D-cultured cells treated with cisplatin for 120 h. Quantitative PCR (qPCR) assays for analysis of the mRNA expression levels of (h) p21 and (i) interleukin-6 (indicators of cellular senescent phenotype) in cisplatin-treated cells under 3D cell culture condition. The expression of GAPDH mRNA was used as an endogenous control for the qPCR assays. In all graphs, the results are presented as mean ± SEM from at least two independent experiments performed in duplicate. Student t test (g), one-way analysis of variance (ANOVA) followed by Tukey post-test (h, i) and two-way ANOVA and the Bonferroni post-hoc test (e) were used for statistical analysis and the differences were considered significant for **P ≤ 0.01 and ***P ≤ 0.001

Back to article page