Fig 1. Ectopic expression of TRIM56 inhibits ZIKV-MR766 infection.

(A) HeLa-FitA2-T56 cells were pretreated with or without 1 μg/ml Dox for 36 h, followed by infection by ZIKV MR766 strain at indicated MOIs in the presence or absence of Dox for 72 h. The expression of ZIKV E, HA-TRIM56 and β-actin (loading control) proteins was determined by immunoblotting. T56, TRIM56. (B) HeLa-FitA2-T56 cells were manipulated as described in (A) and the progeny ZIKV yields in culture supernatants were determined by TCID₅₀ assay. Student t-test, *P<0.05, **P<0.01. Results were representative of three independent experiments. (C) Immunostaining of ZIKV E protein (green) in HeLa-FitA2-T56 cells with or without HA-TRIM56 induction by Dox treatment and infected with ZIKV MR766 strain (MOI = 0.5). DAPI was used for counterstaining of nuclei (blue). Images were representative of two independent experiments. (D) HEK293-Fit-T56 cells were manipulated as described for HeLa-FitA2-T56 cells in (A), followed by immunoblotting of ZIKV E, HA-TRIM56, and β-actin (loading control) proteins. (E) HEK293-T3Y cells with or without expression of FH-TRIM56 were infected with ZIKV MR766 strain for 72 h, followed by immunoblot analysis of ZIKV E, FH-TRIM56, and β-actin (loading control) proteins. FH-T56, Flag-HA-TRIM56. Immunoblots in panels A, D, and E were representative of at least three independent experiments.