PDGF+Cre vs. PDGF only tumors; genes with >2-fold downregulated TE and FDR<0.05.

<p>PDGF+Cre vs. PDGF only tumors; genes with >2-fold downregulated TE and FDR<0.05.</p

<i>In vivo</i> quantification of ribosome-bound and total RNA levels revealed a broad range of ribosome recruitment efficiencies amongst mRNA transcripts.

<p>(A) Distribution of mRNA expression in ribosome-bound and total RNA pools from PDGF-driven glioma identified differential TE (N = 4). (B) TE values for each biological replicate (black points) plotted with the average of the other three replicates (red line) demonstrated reproducibility of measurements. (C) Signal-to-noise ratios of TE measurements (blue bars) identified range of high confidence measurements relative to a normal distribution (red line). (D) GSEA identified statistical overrepresentation of defined gene ontologies amongst efficiently and inefficiently translated genes. Black bars represent distribution of mRNAs from indicated geneset amongst all genes ranked by signal to noise ratio (top panel). Red line represents GSEA output enrichment score. R = Pearson correlation coefficient.</p

GBM is a heterogeneous tumor composed of multiple cell types, including both tumor and stromal cells.

<p>(A) Immunohistochemical staining of mouse PDGF-driven glioma identified multiple tumor regions composed of different cell types. (B) PDGF-HA oncogene expressed from tumor cells colocalized with Olig2 staining by IHC. (C) FACS analysis of tumors generated in Olig2-eGFPL10a mice with PDGF-RFP bicistronic retrovirus identified tumor cells as Olig2+. Scale bar, 100 µm.</p

PTEN loss altered the translation state of glioma cells.

<p>(A) TE measurements of PTEN WT and PTEN deleted tumors were very similar (N = 4). (B) Changes in both total RNA and TE contributed to alteration in ribosome-bound RNA caused by PTEN loss. (C) GO genesets associated with oxidative phosphorylation were amongst those most enriched for transcripts translationally downregulated in PTEN-deleted vs PTEN WT tumors. Enrichment plots for electron transport chain and cellular respiration genesets shown. (D) Genesets associated with glycolysis predominated amongst GO genesets most enriched for transcripts translationally upregulated in PTEN-deleted vs PTEN WT tumors. Enrichment plots shown for genesets for hydrolase activity acting on glycosyl bonds and transferase activity transferring glycosyl bonds. NES = Normalized Enrichment Score. R = Pearson correlation coefficient.</p

Translation efficiency was altered in glioma compared to normal brain OPCs.

<p>(A) Changes in ribosome-bound mRNA correlated somewhat with changes in total cellular RNA between tumor and normal brain OPCs (N = 4). (B) Alterations in TE between tumor and normal brain were reproducible, exceeding average replicate error. (C) Genesets associated with cell division and biosynthetic pathways predominated amongst GOSlim Biological Process genesets most enriched for transcripts translationally upregulated in Olig2+tumor cells compared to normal brain OPCs. Enrichment plots for chromosome organization and mitosis genesets shown. (D) Genesets associated with synaptic signaling predominated amongst GO Biological Process genesets most enriched for transcripts translationally downregulated in Olig2+tumor cells compared to normal brain OPCs. Enrichment plots for transmission of nerve impulse and synaptic transmission shown. NES = Normalized Enrichment Score. R = Pearson correlation coefficient.</p