FIG 3.

HSV-1 gH/gL and gB interact at a steady level and independently of nectin-1. (a) Interaction assay experimental setup. Cells are transfected with viral proteins required for fusion, including split-luciferase-tagged proteins of interest. Interaction is measured by luminescence before and during fusion. Fusion is induced by the addition of target cells expressing the viral receptor nectin-1. (b) gH/gL and gB interaction over time, with target cells expressing or lacking nectin-1. The Halo-Sm and EBV gH-Lg conditions were negative controls. gB-Sm/gB-Lg and PRKACA/PRKAR2A were positive controls. The shaded regions are the SEM. (c) The interactions are quantified by calculating the area under the curve (AUC). (d and e) Total cellular expression and cell-surface expression of Sm-tagged gB H516P—which locks gB in its prefusion conformation—by Western blotting and flow cytometry, respectively. (f) The interaction between gH/gL and gB H516P. Columns show the mean. Error bars are the SEM. ns, not statistically significant; **, P < 0.01. Data in all panels are three biological replicates from independent experiments. Diagrams were created with BioRender.com.