FIG. 5.

LARGE-derived glycan modifications but not O-mannosyl glycans are the crucial structures recognized by arenaviruses. (A) Binding of laminin and viruses to α-DG derived from Lec15.2 cells and wild-type CHO cells with or without overexpression of LARGE. O-mannosylation-deficient Lec15.2 and wild-type CHO cells were transfected with recombinant AdVs expressing LARGE or EGFP. After 48 h, membrane lysates were subjected to WGA affinity purification, and eluted proteins were separated by SDS-PAGE and probed with mAb 8D5 to β-DG in Western blots as described in the legend of Fig. 2A. α-DG was tested for the binding of laminin, LFV, Mobala virus, Oliveros virus, and LCMV as described in the legend of Fig. 3D. (B) Rescue of LFV and LCMV infection in Lec15.2 cells. Lec15.2 and wild-type CHO cells were plated in 96-well plates and infected with AdV-LARGE or AdV-EGFP. After 24 h, cells were infected with pseudotypes of LFV, LCMV, and VSV (MOI of 1), and infection was assessed after 48 h as described in the legend of Fig. 2F.