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. Overall design: mRNA profiles of BMDMs transfected with control ASO or Acly-E14 ASO were generated by deep sequencing.