Fig. 5: Loss of KDM4C leads to differential methylation of target histones and induction of cellular senescence.
From: Histone demethylase KDM4C is a functional dependency in JAK2-mutated neoplasms
A Western blot analysis in BaF3/JAK2-V617F_Cas9_Blast, SET-2_Cas9_Blast and HEL_Cas9_Blast cells following CRISPR-Cas9 knockout using KDM4C specific sgRNAs (KDM4C-sg2, -sg3) or non-targeting control (LUC). B Immunofluorescence analysis of H3K36me3 and Hoechst-staining in SET-2_Cas9_Blast and HEL_Cas9_Blast cells following CRISPR-Cas9 knockout using KDM4C specific sgRNAs (KDM4C-sg2, -sg3) or non-targeting control (LUC). C GSEA showing enrichment of genes related to senescence and autophagy in cancer. Plotted are normalized enrichment scores (NES). D Heat map of differentially expressed genes in KDM4C deleted HEL cells versus non-targeting control (LUC); n = 4. Red zones represent higher gene expression (upregulation), blue zones represent lower gene expression (downregulation). E Representative cytospins of SA-beta-Gal-staining of KDM4C depleted HEL cells compared to non-targeting control (sgLUC). Cells exposed to daunorubicin (Dauno) serve as positive control for SA-beta-Gal staining. F Bar graph depicting quantification of SA-bet-Gal staining as depicted in E. n = 3 independent replicates, two-tailed t-test.
