Fig. 7: Myt1 downregulation contributed to the hyperthermia-induced CDK1 hyperactivation and the synthetic lethality with WEE1 inhibition.

a Representative western blot results in five primary HGSOC samples across five independent experiments. HGSOC cells were incubated at hyperthermia (HT, 42 °C) for 90 min, then allowed to recover at 37 °C for the indicated times (0, 0.5, 1.5, 3, 6, 12, 24, and 48 h). b Representative western blot images of Myt1, CDK1-pY15, and CDK1-pT14 were obtained in ID8 and OVCAR8 cells (Mock) or cells transfected with lentiviral vectors carrying either scramble overexpression (NC) or Myt1 overexpression (OE). c Representative western blot images of CDK1-pY15, CDK1-pT14, and γH2AX proteins were obtained in ID8/OVCAR8 cells transfected with NC or Myt1-OE lentiviral vectors after exposure to either 37 °C or 42 °C for 90 min. d Cell viability was assessed in ID8/OVCAR8 cells (Mock), NC, and Myt1-OE cells following a 90 min HT exposure (n = 14 per group). e EdU and PI staining were analyzed by flow cytometry in ID8/OVCAR8 cells transfected with either NC or Myt1-OE and treated as in (c) (n = 3 per group). f Representative images and quantification of BrdU and γH2AX positive staining in ID8/OVCAR8 cells transfected with NC or Myt1-OE and treated as in (c) (n = 3 per group). Scale bar, 25 μm. g Dose-response curves of cell viability in ID8 and ID8R cells treated with increasing doses of AZD1775 for 48 h (n = 3 per group). h Representative western blot images of the indicated proteins in ID8 and ID8R cells. i Representative western blot images of Myt1, CDK1-pY15, and CDK1-pT14 in cells treated as indicated. Cell viability, measured by CCK8, in cells treated as indicated (j: n = 4 per group; k: n = 3 per group). l Representative western blot images for the indicated proteins were obtained from ID8R cells treated with AZD1775 (800 nM) for 48 h, either alone or in combination with HT. m Dose-response curves were plotted for ID8R cells treated as indicated (n = 3 per group). Comparisons were performed by one-way ANOVA followed by Tukey’s multiple comparisons test in (d, f). Data are presented as mean ± SEM. ****p < 0.0001, ***p < 0.001, ns., not significant. All analytical data are derived from a minimum of three biologically independent experiments, with “n” indicating the specific number of replicates. Source data are provided as a Source Data file.