Fig. 2. MAVI1 negatively regulates RLR-mediated antiviral immune responses in vitro.

(A) Hela cells were infected with lentivirus expressing control shRNA (shCTL) or two individual shRNAs specifically targeting MVAI1 (shMVAI1#1 and shMVAI1#2) and treated with or without poly (I:C) (5 μg/ml, 6 hours), followed by immunoblotting (IB). (B) Hela cells were infected with lentivirus expressing shCTL, shMVAI1#1, or shMVAI1#2 and treated with or without poly (I:C) (5 μg/ml, 6 hours), followed by examination of IFNB1(left) and MAVI1 (right) expression (mean ± SEM, ***P < 0.001). (C) HeLa cells infected with lentivirus expressing empty vector, WT MAVI1 (Flag-MAVI1), or mutant MAVI1 with the first ATG mutated [Flag-MAVI1 (M1)] were subjected to IB analysis. (D) Cells as described in (C) were subjected to examination of IFNB1 expression (mean ± SEM, *P < 0.05, ***P < 0.001). (E) Hela cells were infected with lentivirus expressing shCTL, shMVAI1#1, shMVAI1#2 and treated with or without VSV (2 × 10⁶ p.f.u., 12 hours), followed by examination of IFNB1, MAVI1, and VSV expression (mean ± SEM, **P < 0.01, ***P < 0.001). (F) Hela cells transfected with siCTL or siMAVI1 were treated with VSV-GFP for 12 hours, followed by microscopy imaging analysis. (G) The number of GFP-positive (GFP⁺) cells as shown in (F) were analyzed by ImageJ and normalized to siCTL-transfected cells.