Figure 4.

E3 ligase activity is required for TRIM41 anti-VSV function. (A) HEK293 cells were transfected with FLAG-tagged TRIM41 or the RING deletion mutant (Del-RING). After 24 h, cells were infected with designated MOIs of VSV for 12 h. Relative VSV activities were determined by the luciferase activities that were normalized to the control. Data represent means ± s.d. of three independent experiments. The P value was calculated by two-tailed Student’s t-test. An asterisk indicates p < 0.05. (B) TRIM41 knockout HEK293 cells were transfected with pCMV-3Tag-8 vector, TRIM41-FLAG, or the Del-RING for 24 h, then infected with 0.1 MOI of VSV-GFP for 12 h. The percentage of GFP-positive cells was adopted to determine infection activity. An asterisk indicates p < 0.05. (C) TRIM41 HEK293 knockout cells were transfected with wild type TRIM41 or the Del-RING mutant for 24 h, followed by infection of 0.001 MOI of VSV. After the designated hour post-infection, virus titers were determined by TCID₅₀ in Vero cells. All experiments were biologically repeated three times. The p value was calculated (two-tailed Student’s t-test) by comparison with the wild type. An asterisk indicates p < 0.05.