FIGURE 2 from Epigenetic Priming by Hypomethylation Enhances the Immunogenic Potential of Tolinapant in T-cell Lymphoma

Measurement of gene expression changes in TCL cell lines after treatment with decitabine in vitro. A, Western blots of human (H9 and Karpas-299) and mouse (BW5147 and CT26) cell lysates prepared after treatment with decitabine for 4 days (human) or 2 days (mouse). Dotted lines indicate a cropped blot for presentation purposes. B, Bisulfite pyrosequencing data from four human TCL cell lines, showing basal (untreated) levels RIPK3 promoter methylation (top left), decitabine concentration-dependent decrease of RIPK3 methylation in all cell lines after 4 days treatment (top right), time dependent of RIPK3 promoter demethylation in Karpas-299 cells (bottom left), LINE-1 demethylation as control (at day 4, bottom right) for all four cell lines. Relative levels if cDNA expression measured by qPCR prepared from Karpas-299 (C) and H9 (D) cells treated for 4 days with a various concentration of decitabine. Each gene was normalized to 18S housekeeping gene as a control. * Indicates an estimation of relative expression since this gene was not detected in DMSO control (DMSO control CT set to 40).