Figure 9.

E. chaffeensis infection requires RAB5, and GFP-RAB5 traffics to the E. chaffeensis inclusion membrane. (A) E. chaffeensis infection requires RAB5. RF/6A cells were transfected with control scrambled siRNA (Neg.) or RAB5A/B/C siRNAs for 1 d, and then infected with E. chaffeensis for 2 d. Western blotting was performed using anti-P28, -ACT/actin, or -RAB5 IgG. The values under the bands show the relative ratio of band intensities normalized against ACT/actin, with the ratios of those from control siRNA set as 1. (B to D) E. chaffeensis-infected RF/6A cells at 1 d p.i. were transfected with GFP-RAB5A, GFP-RAB5B, or GFP-RAB5C. At 15 h p.t. (39 h p.i.), cells were subjected to immunofluorescence labeling with rabbit anti-P28 (AF555). Merged, merged images; Merged/DIC, fluorescence image merged with DIC image. Each boxed area is enlarged on the right. (E) Control uninfected RF/6A cells transfected with GFP-RAB5A. Scale bars: 15 μm.