Next Generation Sequencing Facilitates Quantitative Analysis of Bone Marrow-Derived Macrophages Transcriptomes for Acly-E14 ASO effects

Purpose: The purpose of this study is to detect activated or silenced genes during bone marrow derived macrophages (BMDMs) transfected with control siRNA or Acly-E14 siRNA. 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. BMDMs were transfected with control ASO or Acly-E14 ASO. 48 hour later, they were stimulated with LPS (100ng/ml) for 4 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 variation between BMDMs transfected with the indicated ASOs. mRNA profiles of BMDMs transfected with control ASO or Acly-E14 ASO were generated by deep sequencing.