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. 2022 Feb 9;7:37. doi: 10.1038/s41392-021-00857-0

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — SOX2 decreases TET2 expression and 5hmC levels in GSCs and correlates with loss of TET2 in GBM. a Kaplan–Meier survival curves comparing GBM patients across multiple datasets. Survival data were retrieved from the GlioVis portal (http://gliovis.bioinfo.cnio.es). We analyzed datasets that included more than 100 patients. Survival analysis was restricted to patients with primary, IDH1/2 wild-type GBM. b qRT-PCR analysis showing decreased TET2 mRNA in low-passage primary GSC isolates. c RNA-Seq data comparing TET2 expression in neural stem cells (NSCs) and glioma stem cells (GSCs). d Violin plot showing the expression of SOX2 and TET2 across five cell clusters. The mRNA level is shown on y-axis as log2 expression and the x-axis represents the different clusters. scRNA-Seq expression data were retrieved and analyzed with BBrowser (v 2.44.4). e Normalized enrichment scores (NES) from gene-set enrichment analysis showing clusters 1, 2, and 4 are enriched for embryonic stem cell (ESCs) signatures. f qRT-PCR analysis showing the selective decrease in TET2 mRNA and increase in DNMT3A mRNA in GSCs expressing exogenous SOX2. g Western blots showing decreased TET2 protein and no change in DNMT3A protein after transgenic SOX2 expression. h Dot blot assay showing global increase in 5mC and reduced 5hmC after transgenic SOX2 expression in GSCs. i qRT-PCR analysis showing increased TET2 mRNA and no change in TET1 mRNA following forced differentiation of GSCs. Dot blot analysis of genomic DNA isolated from GSCs showing increased 5hmC and decreased 5mC after forced differentiation (inset). Statistical significance was calculated using Student’s t-test b, c, f, and i and data are presented as mean ± SD. *p < 0.05