Intrinsic features of Zika Virus non-structural proteins NS2A and NS4A in the regulation of viral replication
Fig 9
Aborting NS4A protein expression rescued ZIKV RNA replication and protein expression.
(a & c) RNA replication of ZIKV E and HA-NS4A in HMC3 (Fig 9A) or U251 (Fig 9C) cells expressing HA-NS4A or NS4A-X. (b & d) Protein expression of ZIKV E and HA-NS4A in HMC3 (Fig 9B) or U251 (Fig 9D) cells expressingHA-NS4A or NS4A-X. Cells expressing HA-NS4A or NS4A-X were infected with 0.1 MOI of ZIKV/SZ01 for 24 or 48 h. Viral RNA copies and HA-NS4A in the cell lysates were determined by TB Green qRT-PCR. Protein expression of ZIKV E and HA- NS4A were detected by western blot analyses. NS4A-X: The cells were transfected with plasmids expressing HA-NS4A, which has a stop codon introduced to abort the translation of NS4A. The differences between groups of control and HA- NS4A or NS4A-X were evaluated by two-tailed Student’s t test. Data are means ± SEM triplicate experiments; *, P< 0.05; **, P< 0.01; ***, p < 0.001.