Fig. 7: Nuclear MTHFD2 catalytic activity is required for mitosis progression.

a Proportion of HCT116 mitotic cells treated with the indicated concentrations of TH9619 inhibitor for 96 hours normalized to DMSO condition; means + s.d. (n = 3), one-sample two-tailed t-test. b Percentage of HCT116 cells treated with DMSO or 63 nM TH9619 inhibitor for 96 hours at different mitotic phases; means + s.d. (n = 3), a minimum of 300 mitotic cells per replicate were analyzed, unpaired two-tailed t-test. c Quantification of anaphase bridges in HCT116 cells treated with DMSO or 63 nM TH9619 inhibitor for 96 hours; means + s.d. (n = 3), a minimum of 40 anaphase cells per replicate were analyzed, unpaired two-tailed t-test. d Representative images of anaphase defects in HCT116 cells treated with DMSO or 63 nM TH9619 inhibitor for 96 hours. DAPI is shown in gray and H3 phospho-Ser10 (H3PS10) is shown in red (second row) or ICA (third row); non-confocal mode, scale bar 10 μm. e Quantification (left) and representative images (right) of micronuclei in HCT116 cells treated with DMSO or 63 nM TH9619 inhibitor for 96 hours. CREST is shown in magenta and DAPI in gray; non-confocal mode, scale bar 10 μm. For the quantification, means + s.d. (n = 3), a minimum of 3500 cells per replicate were analyzed, unpaired two-tailed t-test. f Comparison of the log₂ mean intensity of nuclear levels of H4K20me1 in HCT116 cells treated with DMSO or the indicated concentrations of TH9619 inhibitor for 96 hours; unpaired two-tailed Wilcoxon test. Representative images are shown on the right; non-confocal mode, scale bar 10 μm. Source data are provided as a Source Data file. The n indicates the number of biological sample replicates for a–c, e; and the number of individual cells for f, where 3 biological replicates were pooled.