Fig. 4

DCZ3301 induced mitotic catastrophe. (a) After treatment with DCZ3301 for 48 h, the comet assay was carried out to evaluate the degree of DNA damage. Scale bars = 100 μm. (b) Immunofluorescence staining of γ-H2A.X in BTZ-resistant NCI-H929R and RPMI-8226R5 cells after treatment with DCZ3301 (8 μM) for 48 h. Scale bars = 5 μm. (c) DCZ3301 activated the ATM-ATR/CHK1 signaling pathway. (d) Western blot analysis showing DCZ3301-mediated activation of the ATM/CHK1 signaling pathway. (e) HR and NHEJ reporter assays of HR repair efficiency and NHEJ repair efficiency in HEK-293 T cells treated with DCZ3301. (f) Quantification of HR and NHEJ repair efficiency. Data are presented as mean ± SD. *p < 0.05, #p > 0.05. (g) Expression of DNA repair related proteins in NCI-H929R and RPMI-8226R5 treated with DCZ3301 for 48 h. (h) Mitotic catastrophe induced by DCZ3301. DNA was stained with DAPI (blue) and the spindles were stained with α-tubulin (red). Scale bars = 5 μm