Fig. 5: IDH1R132H mouse ES cells show replication stress.
From: Mutations inhibiting KDM4B drive ALT activation in ATRX-mutated glioblastomas
a Western blot analyses of WT and IDH1R132H ES cells using antibodies against IDH1R132H, H3K9me3, H3, H3K36me3 and ACTIN. b IP-qPCR analyses of BrdU incorporation at telomeres and Gapdh promoter in WT and IDH1R132H ES cells (n = 3 independent experiments). Immunofluorescence analyses of TERF1 (red; diluted at 1/500) and γH2AX (green; diluted at 1/1500) in WT and IDH1R132H ES cells with and without 1 mM APH treatment for 5 h. Arrows indicate co-staining of γH2AX and TERF1 (c). Percentages of co-localised TERF1 and γH2AX foci are shown in d. For each cell line, >1500 telomeric or TERF1 foci were counted (n = 4 independent experiments). Scale bars: 5 μm. Percentage of co-localised TERF1/γH2AX foci are determined as percentages of telomeric foci that co-stained with γH2AX over the total number of telomeric foci counted. b, d Data are presented as mean values ± SD. **p < 0.005, Student’s t-test with two-tailed distribution. a, c Representative images and blots from at least n = 3 independent experiments. Source data are provided with this paper.
