PML::RARA_WT-V5_anti-V5_CUTandRUN_replicate_2
Bone marrow cells from C57Bl/6 mice were lineage-depleted and retrovirally transduced with murine stem cell virus (MSCV) vectors containing an internal ribosome entry site (IRES) followed by a Thy1.1 cDNA to allow for the purification of transduced cells. These vectors either contained no insert ("empty vector"), WT PML::RARA cDNA with a V5 epitope tag on C-terminus (PR-V5 or PML::RARA_WT-V5), or a V5-tagged C88A mutant in the RARA domain of PML::RARA (C88A or PML::RARA_C88A-V5). Two days following the transduction transduced cells were purified with anti-Thy1.1 magnetic bead enrichment, treated with 1 mM diisopropyl fluorophosphate, and CUT&RUN was performed with anti-V5 or isotype control (IgG) antibodies.