Intrinsic features of Zika Virus non-structural proteins NS2A and NS4A in the regulation of viral replication
Fig 7
The ISGF3 signaling pathway was partially responsible for the antiviral effect of NS4A.
(a) Effect of Abrocitinib on the expression of ZIKV E protein in HMC3 cells. (b) Effect of Abrocitinib on the reproduction of ZIKV in HMC3 cells. HMC3 cells expressing NS4A were infected with 0.1 MOI of ZIKV/SZ01, with (1 μM or 5 μM) or without the treatment of Abrocitinib. Protein expression of STAT1, p-STAT1 (Tyr701), STAT2, p-STAT2 (Tyr690), IRF9 and ZIKV E was detected by western blot analyses (Fig 7A) and viral RNA copies in the culture supernatants were determined by TaqMan qRT-PCR at 48 h p.i. (c) Effect of knockdown of STAT2 on ZIKV E protein expression in U251 cells. (d) Effect of knockdown of STAT2 on ZIKV E RNA replication in U251 cells. The STAT2 in U251 cells with or without NS4A expression was knocked down by shSTAT2. Subsequently, the above cells and the control cells were infected with 0.1 MOI of ZIKV /SZ01. Protein expression of STAT1, p-STAT1 (Tyr701), STAT2, p-STAT2 (Tyr690), IRF9 and ZIKV E was detected by western blot analyses (Fig 7C) and viral RNA in the cell lysates was determined by TB Green qRT-PCR (Fig 7D). The differences of ZIKV viral RNA copies between groups of control and Abrocitinib or shSTAT2 were evaluated by two-tailed Student’s t test. Data are means ± SEM of triplicate experiments; *, P< 0.05; ***, p < 0.001.