Fig. 5

Chemical inhibition of CDK12/13 induces an irreparable DNA damage of MYC-driven Group 3 MB cells. A Workflow scheme of the experiment whose results are shown in B-E. B-E WB (B,D) and densitometric (C,E,) analysis of DAOY and ONS-76 (B,C), D283, MED-411, D341 and HD-MBO3 (D,E) cell lines treated with 100 nM THZ531 for 8 h (T) or DMSO (C), washed and cultured for another 24 h after removal of the treatment (R). HSP90 was used as loading control. Blots in B and D are representative of 3 independent experiments. Graphs in C and E show the mean ± SD of 3 independent experiments. Statistical analysis was performed by One-Way ANOVA test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). F-G Comet assay (F) and relative quantitative analysis (G) performed in MB cells treated or not with 100 nM THZ351 for 8 h after which the drug was carefully removed and cells were left to recover for the indicated time. Representative images in F are shown from 3 independent experiments. Scale bar: 10 μm. Graph in (G) shows data presented as tail moment ± SEM of 3 independent experiments. Statistical analysis was performed by One-Way ANOVA test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)