Fig 2: pUL56 Modulates Immune Receptors through Control of Host Trafficking to the Plasma Membrane(A) Schematic of the experimental workflow. SILAC-labeled cells were infected at an MOI of 10 or mock infected. Samples were harvested 6 hpi and processed for plasma membrane enrichment and subsequent quantitative MS. Data shown in Tables S6 and S7.(B) Hierarchical cluster analysis of fold-change values for each pairwise comparison. Proteins were included if they were annotated as plasma membrane (PM), cell surface (CS), or extracellular (XC) by Gene Ontology (GO), or with a short GO (ShG) term as previously described (Weekes et al., 2014). An enlargement of three clusters is shown in the right panel, which included proteins downregulated during infection with HSV-1 WT but rescued by infection with HSV-1 ?UL56.(C) Profiles of example proteins that were downregulated >2-fold by HSV-1 WT and rescued >2-fold by HSV-1 ?UL56 are shown, as well as the controls gB (expressed only in infected cells), NECTIN-1 (removed from the cell surface by HSV-1), and EGFR (unchanged).(D) U2-OS cells were transfected with FLAG-TLR2 together with pUL56 or pUL56-AAXA expression plasmids. One day post-transfection, cells were stained for surface TLR2 before fixation or intracellular TLR2 after fixation and permeabilization (both red) and co-stained for pUL56 (green). The merge includes DAPI (blue). The scale bars represent 10 µm.