Fig. 1.

The RRV E2 N-linked glycans contribute to MBL deposition onto infected cells. (A) C2C12 myotubes were infected with either RRV WT or E2 DM, and incubated in WT or MBL-DKO mouse serum. Western blots of MBL-C deposition onto cells are shown (left) and viral titer (right) (B) Same as above, but primary C57BL/6 myotube cultures were infected with either RRV WT or E2 DM. Western blots of MBL-C deposition are shown (left) and viral titer (right). (C) C2C12 myotubes were infected with either RRV WT, E2 N200Q, or E2 N262Q and incubated with MBL-WT mouse serum and western blot of MBL-C deposition is shown. (D) Immunofluorescence of either mock (top row), RRV WT (middle row) or RRV E2 DM (bottom row)-infected BHK-21 cells incubated with rhMBL-C at 12 hpi. (i) Individual panels from left: DIC, MBL-C (Texas Red), RRV antigen (FITC), DAPI, and merge. No MBL-C binding was observed in cells incubated without MBL-C (data not shown). (ii) The percentage of infected cells with MBL deposition between RRV WT and E2 DM was determined by visual analysis of four independent fields. * p < 0.05 by Mann-Whitney analysis.