Fosl2 overexpression and Ezh2 inhibition in mouse embryonic fibroblasts

To assess the impact of AP-1 chromatin remodelling, we over-expressed one AP-1 family member, namely Fra2 (encoded by Fosl2), in mouse embryonic fibroblasts (MEFs). To assess the impact of general chromatin depression, we inhibited the activity of PRC2 component histone-lysine N-methyltransferase Ezh2 in MEFS. Overall design: Fosl2 overexpression: We transduced mouse embryonic fibroblasts (MEFs) with a CRISPRa-VPR-mCherry construct and assessed more immediate and longer term effects of Fra2 over-expression by performing OMNI-ATACseq on FACS purified mCherry+/THY1+ cells 4 and 11 days following lentiviral infection with Fra2 gRNAs or scrambled control. For Ezh2 inbition: Following three days of treatment with the Ezh2 inhibitor (Sigma-Aldrich, GSK343) at 4microM and a washout day, MEFs were submitted to the ATAC-seq assay.