FIG. 8.

The major infectious route for MNV-1 is independent of flotillin-1 and GRAF1. RAW 264.7 cells were incubated with an Accell siRNA against flotillin-1 (A), GRAF1 (C), or a nontargeting (NT) construct (A and C) (Dharmacon) for 72 h. Cells were then infected with MNV-1 at an MOI of 10, and VPg expression was determined by immunofluorescence 12 h after infection. To verify flotillin-1 (FLOT) (B) or GRAF1 (D) protein knockdown, protein samples of cells transfected with the respective siRNA construct were analyzed by immunoblotting for flotillin-1 or GRAF1 and quantitated as described in the text.