THP1 cell response to Mycobacterium tuberculosis infection and Mtb PAMP stimulation

To compare gene expression changes induced by infection with Mycobacterium tuberculosis (Mtb) with changes induced by purified Mtb products, we infected THP-1 cells with Mtb strain H37Rv or treated with purified Mtb products, then performed RNAseq. Overall design: THP1 cells were stimulated with PMA for 24 hours, then washed 3 times with PBS, and rested in R10 media for an additional 24 hours. For experiment 1, cells were treated with the indicated PAMPs at 10ug/ml for 8 hours. RNA was then harvested and analyzed by RNAseq. For experiment 2, cells were infected with H37Rv-GFP at an MOI of 10:1 for 4 hours of phagocytosis followed by an 8 hour incubation. Cells were then sorted into infected and bystander populations based on GFP signal. RNAseq libraries were prepared from uninfected, infected, and bystander cells.