Functional analysis of mutations in nsP2 using CHIKV trans-replication system.
U2OS cells were co-transfected with CMV-Fluc-Gluc and CMV-P1234 or plasmids expressing mutant versions of ns polyprotein. Cells co-transfected with CMV-Fluc-Gluc + CMV-P1234GAA served as controls. Cells and media were harvested at 18 h post transfection.(A) Location of mutations in the coding region of CMV-P1234. (B) Expression of ns proteins from CMV-P1234 and its mutant variants (indicated above the graph) was analyzed as described for Fig 3A. (C) Activities of Gluc in the growth media were measured and normalized to those measured for control cells. Data from one out of four reproducible independent experiments is shown.