Fig. 5: hnRNP H/F drive genomic instability and therapy resistance.

a Immunofluorescence experiments in LN18 cells using the γ-H2AX, 53BP1 antibodies and DAPI staining. Mean intensities of γ-H2AX and 53BP1 in 2322 cells were plotted; the bottom and top of the box present the first and third quartile, respectively; the band inside the box shows the mean and the whiskers show the upper and lower extremes. Statistical significance was performed on the full cell populations. n = 2322 cells examined. Shown is a single representative field from one experiment over n = 2 independent experiments. For γ-H2AX: ***P-value = 4.26e-10, for 53BP1: ***P < 2.2e-16 (two-sided Mann & Whitney test). b Western blot analysis of γ-H2AX in LN18 cells treated with dose scale of carboxypyridostatin (cPDS) for 24 h. Shown is a representative result from n = 3 independent experiments. c Quantification of the γ-H2AX levels in LN18 treated with cPDS normalized to GAPDH levels and plotted relatively to the untreated condition. Data are presented as mean values ± SEM of n = 3 independent experiments, P-value = 0.0157 and P-value = 0.0457 for the 2 µM and 10 µM cPDS treatment respectively (two-sided paired t-test). d Quantification of DNA repair kinetics by western blot analysis of γ-H2AX after 4 Gy γ-irradiation in LN18 cells treated with control (siCtr) or hnRNP H/F (siH/F) siRNAs. Shown is a representative result from n = 2 independent experiments. e Plating efficiency assays measuring the cell survival fraction in LN18 treated with siCtr or siF siRNAs and submitted to a radiation dose scale. Data are presented as mean values ± SEM of 6 wells, P-value = 0.0003 and P-value = 0.0006 for the 2 Gy and 4 Gy dose, respectively (two-sided paired t-test). f Quantification of DNA repair kinetics by western blot analysis of γ-H2AX after temozolomide (TMZ) treatment in LN18 cells transfected with an empty plasmid (pICE) or a plasmid expressing Flag-hnRNP H/F. Shown is a representative result from n = 2 independent experiments. For all panels, source data are provided as a Source Data file.