Figure 8

Effect of ERp57 silencing on F-protein levels and SeV replication. (A) IB for SeV-F, ERp57, calnexin and β-actin in WCEs from AGMK cells transfected (48 h) with scramble-RNA (−) or ERp57-siRNA (siERp57) (+), and mock-infected or infected with SeV (3 PFU/cell) for 24 h (top panels). In parallel samples, virus yield was determined at 24 h p.i. by hemagglutinin titration (bottom panel). Data are expressed as percentage of untreated control of quadruplicate samples. Error bars indicate ± S.D. *P < 0.05. (B) IB for Flag-RSV-F, ERp57, calnexin and β-actin in WCEs from A549 cells transfected with scramble-RNA (−) or ERp57-siRNA (siERp57) (+), and, after 24 h, transfected with RSV-F/ORF/C-Flag construct (RSV) or pcDNA3 empty vector (Mock) in the presence of si-ERp57 or scRNA (top panels). RSV-F (Flag) and calnexin protein levels were quantified as described in Methods, normalized to β-actin levels in the same samples, and expressed as percentage of the relative control. Data represent the mean ± S.D. of three independent experiments (lower panels). *P < 0.05. Full-length blots/gels are presented in Supplementary Fig. 14.