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Schematic diagram of the experimental approach. HeLa cells were infected with VACV WR at a multiplicity of 20 PFU/cell. Cycloheximide, harringtonine, or lactimidomycin was added at 2, 4, and 8 h postinfection, and mRNAs protected by ribosomes from digestion with RNase I were isolated and deep sequenced (ribosome profiling). Total mRNA in the presence of cycloheximide was simultaneously isolated, fragmented by digestion with RNase III, and deep sequenced (mRNA profiling).
