Fig. 3: MGMT fusions protect from TMZ-induced damage.

a Clonogenic survival assay of U251 clones expressing MGMT fusions exposed to O6-BG (100 μM) or/and TMZ (100 μM) for 12 days. U251sgMSH6 cells are shown as control for TMZ resistance independently from MGMT. b Cell-cycle distribution of U251 MGMT fusion expressing cells in presence of O6-BG (100 μM) or/and TMZ (100 μM) for 72 h, measured by propidium iodide (PI) staining and FACS. U251 sgCtrl and sgMSH6 are shown as control. c High-throughput microscopy-mediated quantification of cell-cycle distribution at 48 h after treatment. See “Methods” for details. d Quantification of the percentage of cells in c. Data are from a representative experiment repeated in triplicate and presented as mean (technical replicate n = 3) and standard deviation. e, f High-throughput microscopy-mediated quantification of γH2AX intensity levels and 53BP1 foci in U251 cells expressing the MGMT fusions after 48 h of treatment with 100 μM of the indicated drugs. U251 sgCtrl and sgMSH6 were included as controls. The bottom and top of each box represents the first and third quartiles, and the line inside is the median. The whiskers correspond to 1.5 times the interquartile range. Data are representative of n = 3 biologically independent experiments. Two-sided Student’s t test with Bonferroni adjustment for multiple comparisons: ***P < 0.001, **P < 0.01, *P < 0.05, ns not significant, A.U. arbitrary unit. Source data are provided as a Source Data file.