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Fig. 2 | Clinical Epigenetics

Fig. 2

From: Therapeutical interference with the epigenetic landscape of germ cell tumors: a comparative drug study and new mechanistical insights

Fig. 2

Global analysis of the histone modification landscape revealed off-target effects of each epi-drug. A An unsupervised hierarchical clustering heatmap summarizes the results of the mass spectrometry analysis (Mod Spec) of 2102EP cells treated for 16 h with indicated epi-drugs (n = 3). BE The bar charts show the differences in peptide abundancy for histone modifications of the inhibitor target molecules HDAC1 (Quisinostat (B)), KDM5A (JIB-04 (C)), SUV39H1 (Chaetocin (D)) and EZH2 (GSK343 (E)). For raw data, see Additional file 11: Table S1 A (technical replicates n = 3). F Detected changes at indicated histone modifications as found by Mod Spec analysis. G Validation of changes given in (F) using western blot analysis. Data was normalized to total H3 or H4 and densitometrically evaluated. Cells were treated with indicated epi-drugs for 16 h. For raw data, see Additional file 6: Fig. S6F. H Analysis of additional GCT cell lines for changes in the same modifications as given in (FG). For raw data, see Additional file 6: Fig. S6F. I DNA methylation analysis using the slot blot technique. 2102EP cells were treated with indicated epi-drugs for 24 h. A separate membrane was used for each detection. A dilution series (n = 3) was prepared for the three membranes at once. Methylene blue staining served as loading control. J Densitometric analysis of the slot blot membranes. Pixel density of 5mC and 5hmC blots was normalized to methylene blue staining. Changes in DNA methylation are given as percentage of solvent control (set to 100%). Statistical analysis was performed using paired t test (*p < 0.05, **p < 0.005)

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