Fig. 5: MTHFD2 loss impairs mitosis progression.
From: Nuclear localization of MTHFD2 is required for correct mitosis progression
a Percentage of mitotic cells in HCT116 MTHFD2 wild-type (WT) and knock-out (KO1, KO2) conditions, measured with the mitotic marker histone H3 phospho-Ser10 by high-throughput immunofluorescence; means + s.d. (n = 5), unpaired two-tailed t-test. b Percentage of WT, KO1, and KO2 cells in G1 phase at 0, 0.5, 1, 1.5, 2, and 2.5-hour release after RO-3306 drug treatment for 20 hours; means + s.d. (n = 3), at indicated times, unpaired two-tailed t-test. c Percentage of WT, KO1, and KO2 cells at different mitotic phases; means + s.d. (n = 3), a minimum of 150 mitotic cells per replicate were analyzed, unpaired two-tailed t-test. Representative images (d) and quantification (e) of uncongressed chromosomes in metaphase in WT, KO1 and KO2 cells. CREST is shown in ICA (left) or magenta (right) and DAPI in gray; non-confocal mode, scale bar 10 μm. For the quantification, means + s.d. (n = 3), a minimum of 10 metaphase cells were analyzed, unpaired two-tailed t-test. Representative images (f) and quantification (g) of anaphase defects in WT, KO1 and KO2 cells. CREST is shown in ICA (left) or magenta (right) and DAPI in gray; non-confocal mode, scale bar 10 μm. For the quantification, means + s.d. (n = 3), a minimum of 25 anaphase cells were analyzed, unpaired two-tailed t-test. h Difference between the beta score of all genes in KO and WT cells. Synthetic lethal hits with MTHFD2 KO with a beta score < −1 are shown in red, and synthetic viable hits with MTHFD2 KO with a beta score > 1 are shown in blue. Shared hits between both KOs are indicated with a black stroke. i Percentage of survivor WT, KO1, and KO2 cells normalized to DMSO after etoposide treatment with indicated concentrations for 72 hours; means + s.d. (n = 3), unpaired two-tailed t-test. Inside the graph, a representative scanned image of one replicate (4 technical replicates are shown per condition). Source data are provided as a Source Data file. The n indicates the number of biological sample replicates for a–c, e, g, i.
