Mutations in NS5A domain I disrupt the production of infectious virus.
In vitro transcripts of mJFH-1 containing the indicated mutations were electroporated into either Huh7 (A, B), or Huh7.5 (C-E) cells. Virus genome replication and protein expression was assayed by quantification of NS5A positive cells 48 h.p.e. for Huh7 (A) or Huh7.5 (C) cells by using the Incucyte-ZOOM . (B, D) Intracellular and extracellular infectious virus was titrated at 72 h.p.e. E Huh7.5 cell lysates at 72 h.p.e. were analysed by western blot with anti-NS5A, anti-Core and anti-β-actin antibodies. Data from three independent experiments are shown and error bars represent the standard error of the mean.