FIG 7.

T483C/V484C and V484C/N485C bind a prefusion conformation-specific antibody prior to cell disruption. (A) Vero cells were transfected with WT HeV F or one of the TMD mutants. At 24 h posttransfection, the cells were metabolically labeled for 24 h. Then, the cells were treated with HeV F MAb 5B3 antibody for 1 h, followed by lysis and pulldown. Control samples were lysed after the overnight label and treated with HeV F anti-peptide Ab for immunoprecipitation. The samples were analyzed by 10% SDS-PAGE. (B) 5B3 binding was quantified by densitometry and normalized to the level of WT HeV F. All data are represented as the means ± standard deviations for three independent experiments. (C) Vero cells were transfected with WT HeV F or one of the TMD mutants. At 24 h posttransfection, the cells were metabolically labeled for 3 h. Then, the samples were immunoprecipitated with anti-peptide Ab or MAb 5B3 and analyzed by 10% SDS-PAGE.