FIG 2.

Updating the MHV68 intraviral protein interaction network. (A) Viral proteins identified by MS were validated by coimmunoprecipitation assays with mLANA-GFP. 293T cells were transfected with plasmids encoding the indicated FLAG-tagged viral proteins and mLANA-GFP. At 48 h posttransfection, cells were lysed and mLANA-GFP was immunoprecipitated from lysates using GFP antiserum. Immunoblot analyses were performed with FLAG-specific antibodies to detect coprecipitating viral proteins. (B) GST and mLANA-GST were purified from bacterial lysates and incubated with lysates from 293T cells transfected with plasmids encoding mLANA-FLAG, ORF59-FLAG, or ORF57-FLAG. Complexes were captured using glutathione agarose. Precipitates were washed and resolved by SDS-PAGE. Immunoblot analyses were performed to detect FLAG- and GST-tagged proteins. (C) The mLANA intraviral protein interaction network. Blue nodes represent viral interactions identified in this study, yellow nodes represent viral interactions identified previously (54), green nodes indicate interacting viral proteins identified both in this study and previously (54), square nodes designate interactions validated by coimmunoprecipitation in this study, solid red edges designate specific interactions scored by I-DIRT, dashed red edges designate nonspecific scored interactions, dashed purple edges represent unscored interactions, and solid black edges represent interactions validated in previous studies. WCL, whole-cell lysate; IP, immunoprecipitation; IB, immunoblot.