Fig. 7

Genes associated with H4K5acK8ac-preferred super-enhancers are involved in the glioblastoma stem-like properties. A Schematic representation showing the CRISPR-Cas9–mediated genome editing approach for SEs. Guide indicates gRNA. B Deletion of the H4K5acK8ac-preferred SEs associated with MYCN and NFIC in 0316-GSC cells. Expected band sizes of genomic DNA for unedited (arrowhead) and SE-edited samples (asterisk) are marked. Images of the uncropped gel are shown in Figure S9A. C–G Biological effects of deletion of the H4K5acK8ac-preferred SEs associated with MYCN and NFIC in 0316-GSC cells. C Quantitative reverse-transcription PCR analysis of the expression of MYCN and NFIC and non-target genes following SE deletion (n = 3). D Cell proliferation rates at 4 days after SE deletion (n = 4). E Expression of stem cell marker genes, NESTIN (left) and SOX2 (right), at 4 days after SE deletion (n = 3). F Phase-contrast images of 0316-GSC cells at 14 days after SE deletion. Images are representative of three independent experiments. Scale bar, 50 μm. G In vitro sphere formation efficiency of 0316-GSC cells at 2 weeks after SE deletion (n = 3). C–E and G Data are means ± SEM *P < 0.05, **P < 0.01 (two-tailed Student’s t-test)