Fig 2: KDM4C is a key activator of the TGF-ß2/Smad/ATM signaling pathway.a, b Changes in essential components of the TGF-ß signaling pathway after genetical or pharmacological inhibition of KDM4C in SPC-A1 cells (n = 3). c, d SPC-A1 cells stably expressing control or KDM4C shRNAs were exposed to irradiation and collected 4 h later. Immunostaining was performed to examine p-ATM and p-Chk2 foci formation. ***P < 0.001 (n = 3). Scale bar, 10 µm. e, f SPC-A1 cells were treated with SD70 (3 µM) for 24 h and 48 h, respectively prior to exposure to irradiation. Cells were harvested after 4 h and subjected to immunostaining using p-ATM and p-Chk2 antibodies. Foci formation was quantified under a fluorescent microscope. ***P < 0.001 (n = 3). Scale bar, 10 µm.

Fig 3: ALDOA promoted PLK1 activation by suppressing ATM expression. A and B The expression of ATM, phospho-PLK1-T210, PLK1 and ALDOA was determined by western blot analysis. C Dose-dependent effect of KU-55,933 treatment for 16 h on the phospho-PLK1-T210 and PLK1 protein. The expression of phospho-PLK1-T210 and PLK1 in PANC-1 or SW1990 cells treated with 10 ng/ml KU-55,933 over a time gradient. D and F PLK1 exerted significant regulation of the G2/M phase in cell cycle analysis by flow cytometry. E. The ALDOA group and the control group respectively were transfected by siPLK1 and siNC to assess colony formation capacity. G The expression of ALDOA, phospho-PLK1-T210 and PLK1 was determined by western blot analysis

Fig 4: Schematic illustration depicting the function of ALDOA: ALDOA reverses the cell cycle arrest induced by DNA damage by inhibiting ATM and promoting PLK1 activation. The dashed arrow highlights the alleviation of cell cycle arrest in G2/M phase by phospho-PLK1-T210 in tumour cells without ALDOA

Fig 5: ALDOA decreased the percentage of cells in G2/M phase by suppressing ATM expression. A GSEA was conducted to analyse the signalling pathway enrichment in siALDOA PANC-1 and SW1990 cells compared with the NC group. B, C, D and E The mRNA and protein levels of ATM expression in ALDOA-silenced and ALDOA-overexpressing cell lines (PANC-1 and SW1990). F Confocal microscopy showed DNA damage (green) and nuclear DNA (blue) in ALDOA-silenced or normal cells (PANC-1 and SW1990) treated with siATM or siNC. G Cell cycle analysis by flow cytometry demonstrated an obvious change in the proportion of cells in G2/M phase in ALDOA-silenced or normal cells (PANC-1 and SW1990) treated with siATM or siNC