FIGURE 10:
HSV-1 infection mobilizes M6P, with destruction and depletion of LLOs. (A) Uninfected PERK−/− MEFs were treated with 2 mM DTT and/or 0.2 mM CHX for 1 h, as indicated, and G3M9Gn2-P-P-Dol was measured (n = 3, mean ± SEM). *, p < 0.0001 for untreated controls compared with DTT-treated samples; #, p < 0.002 for the latter compared with DTT + CHX-treated samples. (B) Normal (PERK+/+) MEFs were stressed with TG (gray) or infected with HSV-1 (black). G6P, M6P, G3M9Gn2-P-P-Dol LLO, and free glycans in the G4–G7 range (F.G.) are expressed as percentages of untreated cells (white), average of duplicates ± range. Cells were infected at multiplicity of infection (moi) = 7.5 and analyzed 12 h.p.i. (C) FACE gel showing free glycans associated with (B). (D) PERK+/+ MEFs were infected as for (B). Total hexose-1-Ps (white squares), total hexose-6-Ps (white circles), free glycans in the G4 -G7 range (black circles), and G3M9Gn2-P-P-Dol (black squares) are expressed as percentages of 5 h mock infection, averages of duplicates ± range (error bars are obscured by symbols).