Fig. 2.

Detection of phosphorylated ribosome-associated proteins during viral infections. (A) Silver stain of GFP-TRAP complexes from uninfected or VacV-, HSV-1- or VSV-infected (MOI=5) HAP1 cells expressing RACK1–eGFP at 20 hpi (for VacV and HSV-1) or 6 hpi (for VSV). (B) Venn diagram of ribosome-associated proteins identified by mass spectrometry of GFP-TRAP complexes as being uniquely phosphorylated in one or more infected sample, but not in uninfected samples. (C,D) Western blot analysis of samples from HAP1 cells depleted of RPS2 or RPS28 using two independent siRNAs (Si-1, Si-2) following infection with VacV (MOI=0.5) for 20 hpi (n=3). D8 and A14 are poxvirus proteins used to measure infection levels. Heat-shock protein HSP90 serves as a loading control.