Next generation sequence data to elucidate the role of CHD8 in terminal erythropoiesis

To uncover how CHD8 regulates terminal erythropoiesis, we performed CUT&RUN assays to check the direct binding of CHD8 on the target genes in terminal erythroblasts. In addition, we perfromed RNA sequence in sorted ProEs from WT, P53 KO and Chd8 P53 DKO mice to identify the differentially expressed genes due to loss of CHD8. For CHD8 CUT&RUN assay, wild type CD44+TER119+ erythroblasts were sorted out from bone marrow and used for CHD8 binding, followed by Mnase binding and released DNA extraction. DNA were further used for library preperation with NEBNext DNA library preperation kit and send for sequencing. In analysis, after mapping, two replicates were merged into one and used for peak calling. For ProE RNA sequencing, WT, P53 KO and Chd8,P53 DKO mice bone marrow were harvested at 2 weeks post pI:pC induction and ProEs were sorted out by BD sorter. The ProEs were lysed with Trizol and RNA extracted with Qiagen microRNeasy Kit. The RNAs were send to the core for library preperation and sequencing.