FIG 6.

HeV G stalk N-glycan mutants modulate receptor-induced conformational changes differently than WT HeV G. (A) Ephrin B2-deficient PK13 cells were transfected with WT or mutant HeV G to assess cell surface expression (CSE) and MAb26 and MAb45 binding in the absence of receptor by flow cytometry. The CSE, MAb26, and MAb45 binding levels of the HeV G N-glycan mutants were normalized to the corresponding levels obtained for WT HeV G (set to 100%). (B) The ratios of MAb26 or MAb45 binding levels to CSE were determined and normalized to that of WT HeV G (set to 100%; dotted line). (C) MAb26 and MAb45 binding levels to HeV G N-glycan mutants were determined in PK13 cells in the absence (0 nM) and presence (100 nM) of soluble ephrin B2. Binding levels in the presence of ephrin B2 were normalized to binding levels in the absence of ephrin B2 (set to 100%; dotted line). The data represent the average results ± SEM from at least three independent experiments.