Fig 9.

JEV infection requires trafficking through Rab5-positive compartments but is independent of Rab7. (A) DiD-labeled JEV (pseudocolored in green) was added to Neuro2a cells transfected with dsRed-Rab5 or GFP-Rab7 (pseudocolored in red) on ice for 1 h. Cells were warmed to 37°C for the indicated times, washed with low-pH buffer, fixed, and imaged. DiD-labeled JEV colocalization with Rab5 is seen only by 30 min postinternalization (lower left panel, arrowheads). Bar, 10 μm. (B) Neuro2a and Vero cells were pretreated with 100 nM bafilomycin before infection with JEV. Infection was quantified as described in Materials and Methods. (C) Neuro2a and Vero cells transfected with GFP, GFP-wt Rab5, GFP-Rab5dn, GFP-wt Rab7, or GFP-Rab7dn were infected with JEV and processed at 24 hpi as described above. Infection was normalized to cells transfected with GFP alone. Student's t test was used to generate P values. **, P < 0.01.