Article Title: Efficient induction of differentiation and growth inhibition in IDH1 mutant glioma cells by the DNMT Inhibitor Decitabine
Figure Lengend Snippet: Decitabine reverses genome-wide DNA methylation and induces expression of genes associated with differentiation in IDH mutant glioma cells A, Low dose DAC inhibits DNMT1. Results from western blot shown. B, DAC treatment results in loss of DNA methylation. DNA methylome analysis of TS603 and TS667 cells following DAC treatment (200nM) is shown. Results from the Illumina HumanMethylation450 array. C, Gene expression changes following DAC treatment (200nM). Results from Affymetrix gene expression arrays. Most significantly altered genes following 200nM DAC are shown. D, Significant concordance between demethylated and upregulated genes and polycomb targets. Venn diagram showing overlap between the gene sets. P value (hypergeometric) is shown.
Article Snippet: Proteins were separated by SDS–PAGE, transferred to PVDF membrane (Millipore) and probed with the following primary antibodies: anti-IDH1 R132H (Dianova, DIA-H09), anti-DNMT1 (New England Biolabs, M0230S), anti-GFAP (Cell Signaling Tech, 2118S) and anti-β-actin (Sigma, A5316).
Techniques: Genome Wide, DNA Methylation Assay, Expressing, Mutagenesis, Western Blot