FIG 9.

MxB inhibits dengue virus and Japanese encephalitis virus. (A) Vero cells were transiently transfected with MxB-Flag DNA and then infected with JEV (SA14-14-2) at an MOI of 0.01. After 48 h, levels of JEV RNA in the infected cells were determined by qRT-PCR. Expression of MxB-Flag was examined by Western blotting. (B) Zika virus (PLCal_ZV) (MOI = 0.5) was used to infect Vero cells transiently expressing MxB-Flag. At 48 hpi, the level of Zika virus RNA in the infected cells was determined by qRT-PCR. (C) Vero cells were transiently transfected with MxB-Flag DNA and then infected with DENV2 (Tr1751) (MOI = 0.5). After 48 h, the supernatants were collected for plaque assay to measure the titers of DENV2. (D) Effects of CsA on replication of HCV, JEV, and ZIKV. HCV (MOI = 0.5), JEV (MOI = 0.01), and ZIKV (MOI = 0.5) were used to infect Huh7.5.1 cells in the presence of different doses of CsA (0.25 μM, 0.5 μM, 1 μM, and 2 μM). After 48 h, levels of viral RNA was determined by qRT-PCR. (E and F) Effects of MxB on expression of NS5. Either JEV NS5-Flag (E) or DENV NS5-Flag (F) was coexpressed with various amounts of MxB-Myc for 24 h, followed by treatment with MG132. The expression of NS5 and MxB was determined by Western blotting. DMSO, dimethyl sulfoxide. (G) MxB interacts with NS5A(DEYN), JEV NS5, and DENV NS5. Immunoprecipitation was performed with anti-Flag antibody to pull down Flag-tagged NS5A(DEYN), JEV NS5, and DENV NS5. Levels of coprecipitated MxB-Myc were determined by Western blotting.