Fig. 1: H3.3G34R promotes ALT in APT-tKO mouse ES cells.
From: Mutations inhibiting KDM4B drive ALT activation in ATRX-mutated glioblastomas
a Glioblastoma tumours from patients aged 30 years and younger with ATRX, H3.3, IDH1 and TP53 mutations. b, c Telomere length analyses of APT-tKO #1 and H3.3G34RAPT-tKO #1 ES cells, in relative to WT (wildtype) ES cells (n = 3 independent experiments). d Detection of telomeric C-circle in H3.3G34RAPT-tKO cells (n = 5 independent experiments). e Detection of T-SCE in H3.3G34RAPT-tKO cells (n = 3 independent experiments). Human U2OS osteosarcoma ALT cell line was included as a positive (+ve) control. f, g Immunostaining of APBs in APT-tKO cells and H3.3G34RAPT-tKO, shown by co-staining of TERF1 (green; diluted at 1/500) and PML (red; diluted at 1/800) (arrows). Scale bars: 5 μm; representative images from at least n = 3 independent experiments. Percentages of co-localised TERF1 and PML foci in H3.3G34RAPT-tKO#1 cells are shown in g. For each cell line, >1500 telomeric foci were counted (n = 3 independent experiments; indicated by TERF1 staining). Percentage of co-localised TERF1/PML foci are determined as percentages of telomeric foci that co-stained with PML over the total number of telomeric foci counted. b, c, d, e, g Data are presented as mean values ± SD. *indicates p < 0.05 and **indicates p < 0.005, Student’s t-test with two-tailed distribution. Source data are provided with this paper.
