Fig. 2.

Trim25 restrains RABV infection in 293T cells. (A–C) 293T cells were transfected with the expression vector pCAGGS or pCAGGS to express the FLAG-Trim25 (FLAG-Trim25) at the indicated volumes. The cells were infected with CVS at an MOI of 0.01 at 12 h post-transfection. After 36 h incubation, the cells were harvested to measure the protein levels of Trim25, RABV-N, and β-actin by Western blotting (A). The supernatants were harvested for virus titration (B). The same treated 293T cells were fixed and stained to measure RABV-N and Trim25 expression using IFA. The merged pictures are shown in the right panels, and the enlarged pictures are placed inside the white dotted box (C). Scale bar, 200 μm. (D, E) 293T cells were transfected with the expression vector pCAGGS or pCAGGS to express FLAG-Trim25 (FLAG-Trim25) at the indicated volumes. At 12 h post-transfection, the cells were infected with a wildtype RABV strain DRV (D) or SHBRV (E) at an MOI of 0.01. The supernatants were harvested for virus titration. Statistical analysis of comparisons between groups was carried out by Student’s t-test (∗, P ​< ​0.05; ∗∗, P ​< ​0.01; ∗∗∗, P ​< ​0.001). The bar graph shows the mean ​± ​SD, n ​= ​3. Western blotting data are representative of at least two independent experiments. See also Fig. S1.