Intrinsic features of Zika Virus non-structural proteins NS2A and NS4A in the regulation of viral replication
Fig 6
NS4A enhanced ISGs expression by activating ISGF3 pathway in HMC3 cells.
(a) Venn Diagram of differentially expressed genes. After HMC3 cells with or without NS4A expression were infected with 0.1 MOI of ZIKV/SZ01 for 24 h, total RNA from the mock and virus infected cells were collected for RNA-Seq, followed by analyses of the differentially expressed genes. Vec-M or Vec-S, mock or ZIKV/SZ01 infected- HMC3 cells without NS4A expression; NS4A-M or NS4A-S, mock or ZIKV/SZ01 infected-group of HMC3 cells with NS4A expression. (b) Expression of ISGs in HMC3 cells expressing NS4A determined by RNA-Seq. (c) Expression of ISGs in HMC3 cells expressing NS4A verified by qRT-PCR. The differences between groups of control and NS4A were evaluated by two-tailed Student’s t test. Data are means ± SEM of triplicate experiments; *, P < 0.05; **, P< 0.01; ***, p < 0.001. (d) The top 20 enriched pathways of Vec-M vs NS4A-M. The pathway enrichment analysis was used to group the differentially expressed genes in HMC3 cells expressing NS4A and vector. (e) Detection of STAT1, p-STAT1 (Tyr701), STAT2, p-STAT2 (Tyr690) and IRF9 protein expression in the ISGF3 signaling pathway in HMC3 cells expressing NS4A by western blot analyses.