Journal: Cell stem cell
Article Title: Adaptive chromatin remodeling drives glioblastoma stem cell plasticity and drug tolerance
Figure Lengend Snippet: Transcriptional Programs Related to Slow Proliferation and Stemness are Enriched in GSC8 Persisters (A) Heatmap shows expression profiles of the 4,084 most variably expressed genes across GSC8 naïve, GSC8 12d , and GSC8 Per . K-means clustering was performed to distinguish sets of genes with coherent patterns of expression across the time course of dasatinib treatment. Data was generated from three biological replicates of separately derived GSC8 12d and GSC8 Per cultures. Color represents z-scores of gene expression across rows. (B) Heatmap shows average changes in gene expression of clusters 1–6 in GSC8, GSC26 and CW1691 after 12-day treatment with gefinitib and dasatinib, respectively, compared to corresponding untreated control. Similar trends of gene expression change are evident in GSC8, GSC26 and CW1691 persisters. Color represents log2(fold-change) gene expression, where the scale ranges from the minimum to maximum value observed in GSC26 12d or CW1691 12d . (C) Gene set enrichment analysis (GSEA) shows enrichment of a cell cycle meta-signature (upper panel), gene sets related to qNSCs (middle panels), and Sox2 + medulloblastoma cells (lower panel) across naïve and persister states for GSC8 and GSC26. The cell cycle gene signature is negatively enriched, while the quiescence gene signatures are positively enriched upon drug treatment. (D) Barplots show the expression levels (reads per kilobase per million mapped reads (RPKM), y-axis) of SOX2, SOX4, OLIG2, NFIA, DLL1, NOTCH1, KDM5A, KDM5B and KDM6B in normal human brain (GTEx), GSC8 naïve, GSC8 12d , GSC26 naïve, GSC26 12d , CW1691 naïve, and CW1691 12d . Most genes are upregulated in drug-treated cells. Error bars represent s.e.m. (E) Barplot shows the fraction of cells (y-axis) CD133 + or CD15 + by flow cytometry. GSC8 Per display significantly increased positivity for CD133 and CD15 in comparison to GSC8 naïve (Student’s t -test; * P
Article Snippet: Notch1 ICD ORF was PCR-amplified from Notch1 intracellular domain-pcw107 (Addgene, plasmid # 64621) using primers containing flanking Gateway attB sites and was subcloned into p-DONR221 and pIND20. dnMAML-GFP ORF was PCR-amplified from MigR1-dnMAML ( ) using primers containing flanking Gateway attB sites and was subcloned into p-DONR221 and pIND20.
Techniques: Expressing, Generated, Derivative Assay, Flow Cytometry