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. 2020 Jul 20;11:3627. doi: 10.1038/s41467-020-17357-4

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© The Author(s) 2020

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. 7 — a–c EIF2 and mTORC1 signaling (highlighted) associated with genes that have significant changes in the H3K4me3 modification, following OTX2 silencing in Group 3 MB tumorspheres. Significance was calculated using a right-tailed Fisher’s exact test. d, e OTX2 silencing in HDMB03 (d) and D283 (e) tumorspheres with two independent siRNAs reduces SOX2, p-S6, p-4E-BP1, and Rheb levels. The neuronal differentiation markers DCX, βIII tubulin, and EPHB2 are increased along with small increases in pRaptor in D283 tumorspheres only. Total S6, total 4E-BP1, total Raptor, GAPDH, and β-actin serve as loading controls. f Enriched biological processes and pathways determined using GSEA (Terms with FDR corrected q-value < 0.25) and visualized in Cytoscape v3.7.2 using Enrichmentmap. Each node in the map denotes a pathway, process or a transcription factor enrichment. The size of the nodes is proportional to the number of genes in the process. The nodes are connected when they share enriched genes. Nodes with asterisk denote the processes enriched with q < 0.05. g Representative images of p-S6 and p-4E-BP1 immunohistochemical staining in three independent FFPE Group 3 (upper) and Group 4 (lower) MB patient samples. Scale = 200 μm.