Ribosome profiling and RNA-seq of WT and TLR4-/- BMDM infected with Salmonella
Ribosome profiling and RNA-seq of WT and TLR4-/- BMDM infected with Salmonella
Macrophages were differentiated from cells harvested from mouse bone marrow. These were then infected with WT or Î”prgJ Salmonella Typhimurium or mock infected. Cells were lysed in buffer containing cycloheximide and chloramphenicol, and lysates were split for RNA-Seq and Ribo-Seq. For Ribo-Seq, lysates were treated with RNase I and fragments protected from digestion by the ribosome were purified. For RNA-Seq, total cellular RNA was fragmented by alkaline hydrolysis. Both produce short RNA fragments, from which libraries were generated.