Ribosome profiling of MCMV

Swiss murine embryonic fibroblasts (NIH-3T3) were infected with wild-type Smith strain mouse cytomegalovirus (MCMV) at a multiplicity of infection (MOI) of 10. Ribosome profiling was performed at various times during infection with minor modification to the protocol described in Stern-Ginossar N et al., Science 2012. The detailed protocol is described in Rutkowski et al, Nature commun 2015, and Whisnant et al., Nature commun. 2020. Translation start site profiling was performed by culturing cells in presence of Harringtonin or Lactimidomycin for 30 min prior to cell harvest. Overall design: Ribosome profiling was performed a 0, 2, 6, 12, 24 and 48 h post infection. Translation start site profiling was performed using Harringtonin (2µg/ml for for 30 min) or Lactimidomycin (50µM for 30 min) prior to cell lysis. Two biological replicates were analysed.