Figure 7.

Late-stage HSV-1 hypoglycosylation is emulated by glucose depletion with LLO loss. LSH was emulated by plating 5 × 10⁵ cells in 12-well plates with 1 ml DMEM plus 10% FBS. Twenty four hours later, cells were infected with HSV-1 at MOI = 1 or MOI = 10, and harvested 24 hpi. A) WT, STT3A-KO, and STT3B-KO cells were cultured with either 5 or 25 mM glucose. gC, gD (with glycoforms indicated), and VP5 were detected by immunoblotting. One set of samples was incubated with 2 μg/ml TN. B) Glucose concentrations in media either prior to infection or 24 hpi, for DMEM formulated with 5 or 25 mM glucose. C) G₃M₉Gn₂-LLO at 24 hpi, normalized to the value for 5 mM glucose medium. D) Glucose consumption by WT HEK cells grown in medium with 5 mM glucose and infected with HSV-1 at MOI = 1 or 10 for 12 or 24 h. Calibration of the assay showed that the limit of sensitivity was 98–99% consumption (i.e., 1–2% glucose remaining); ns, not significant.