Multiple roles of the non-structural protein 3 (nsP3) alphavirus unique domain (AUD) during Chikungunya virus genome replication and transcription
Fig 8
Effect of AUD mutations on CHIKV protein expression and RNA synthesis.
(A) C2C12 cells were electroporated with ICRES-RNAs (wildtype (WT) and indicated AUD mutants) and cell lysates were collected at 36 h.p.e. Expression of nsP3 and capsid was analysed by western blot. Representative western blots are presented. For quantification using a LiCor Odyssey Sa fluorescence imager western blots from three independent experiments were analysed and the graph on the right shows the ratio of capsid to nsP3 expression. (B) C2C12 cells were electroporated with the indicated ICRES RNAs, cellular RNA synthesis was inhibited by actinomycin D and nascent viral RNAs were labelled with [3H]-uridine. A representative autoradiograph is presented. The graph on the right shows the ratio of gRNA to sgRNA determined by quantification of three independent experiments. (C) The same RNAs were fractionated on a sucrose gradient and [3H]-labelled RNAs were detected by scintillation counting of individual fractions.