Chromatin ImmunoPrecipitation DNA-Sequencing after H19 OE and KD
Treatment of pathological cardiac remodeling and subsequent heart failure represents an unmet clinical need. The well conserved lncRNA H19 shows as powerful therapeutic potential in the treatment of pathological cardiac hypertrophy. H19 is strongly repressed in failing hearts from mice, pigs and humans. Gene therapy using murine but also human H19 strongly attenuated heart failure even when cardiac hypertrophy was already established. Using microarray , GSEA and ChIP-Seq we identified a link between H19 and NFAT signalling. H19 physically interacts with PRC2 to epigenetically induced Tescalcin repression which in turn leads to reduced NFAT expression and activity. Overall design: HL-1 cells were transfected with siRNA for H19 KD (50nM: siH19, siCtrl) to induce H19KD or transduced with lentivirus for stable H19 OE (pLV+H19, pLV+empty). Chromatin was isolated and 4 ug of antibody against H3K27me3 were applied for the isolation of genomic DNA regions of interest. ChIP DNA was purified.