Next Generation Sequencing Facilitates Quantitative Analysis of wild type and KAT8-deficient Bone Marrow Derived macrophage Transcriptomes

Purpose: The purpose of this study is to detect activated or silenced genes during wild type (WT) and KAT8-deficient 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. Cells with purity >97.5% were used for subsequent experiments. WT and KAT8-deficient BMDMs were stimulated with R848 (100ng/ml) for 0 and 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 WT and KAT8-deficient BMDMs. Overall design: mRNA profiles of WT and KAT8-deficient BMDMs were generated by deep sequencing.