Figure 3
A) Comparison of fusion activity of wild-type and a fusion peptide mutant of HRSV F. 293T cells co-transfected with pBD-NFκB and either pHRSVFOptA2 or pL138R were mixed 24 hours later with a separate population of 293T cells that had been transfected with pFR-Luc, and luciferase activity was measured 24 hours post mixing as described in methods. Luciferase activity is reported as relative light units. B) Processing of the wild-type and L138R mutant of HRSV F was determined by metabolic labeling 293T cells transfected with either pHRSVFOptA2 (lane 1), pL138R (lane 2), pCMV-β-gal (lane 3), or mock transfected (lane 4) for 6 hours with [35S]-cysteine/methionine followed by immunoprecipitation of lysates with HRSV F specific mAbs, and analysis of immunoprecipitates by SDS-PAGE as described in methods. C) Cell surface expression of the wild-type and L138R mutant F proteins in 293T cells transfected with either pHRSVFOptA2 or pL138R was compared by flow cytometry as described in methods.