Next Generation Sequencing Facilitates Quantitative Analysis of Bone Marrow Derived macrophage administrated with KAT8 inhibitor

Purpose: The purpose of this study is to detect activated or silenced genes following pretreatment with KAT8-inhibitor in bone marrow derived macrophages (BMDMs). Gene expression differences between two samples could be found using transcriptome profiling (RNA-seq) analysis. Methods: Mouse BMDMs were generated from bone marrow cells in RPMI-1640 medium with recombinant mouse M-CSF (20ng/ml). BMDMs were stained to confirm the surface expression of CD11b and F4/80,and the use of BMDMs with purity greater than 97.5% for subsequent experiments.BMDMs pretreated with DMSO or KAT8 inhibitor were stimulated with R848 (100ng/ml) for 2 hours, of which RNA profiles were generated by deep sequencing, using Illumina. Results: We mapped about 10 million sequence reads per sample to the mouse genome, identified hundreds of genes with significant mRNA between DMSO or KAT8 inhibitor-treated BMDMs. Overall design: mRNA profiles of BMDMs pretreatment with DMSO and KAT8 inhibitor were generated by deep sequencing.