![Fig. 3. Pulse-chase experiments of wild-type and mutant FV protein in COS-1 cells. / COS-1 cells, transiently transfected with pMT2/FV (wild type) or pMT2/FV-Arg2074Cys (mutant), were pulse-labeled with [35S]-methionine and [35S]-cysteine for 2 hours, and then chased by cold methionine and cysteine for various periods of time up to 180 minutes. At the specified chase period (0, 30, 60, 120, and 180 minutes) radiolabeled FV was immunoprecipitated from cell lysates (A,B) and from the corresponding conditioned media (C,D), electrophoresed on 4% SDS-PAGE gels under nonreducing conditions, and then detected by a phosphor imager (Typhoon 9200). The arrowheads indicate the 330-kDa FV molecule. In all panels, the pUC18 lane contains immunoprecipitable proteins at the end of the pulse period from COS-1 cells transfected with the unrelated pUC18 plasmid. Below each lane, a densitometric analysis of the band corresponding to immunoprecipitated FV is shown. Bar graphs are expressed as arbitrary densitometry units (y-axis), as calculated by the ImageQuant software by integrating intensities of all the pixels in the band excluding the background.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/101/1/10.1182_blood-2002-06-1928/6/h80133603003.jpeg?Expires=1735559235&Signature=cIJGoLThhgnG7Ym5E-rPrj-UNod2TaWNd3Hj7f408gBhwGKmzhsq2QZKcM2wnqu2HfV01UAeeGE05WaSWj0aLj4lFwwNnB7JmSQpb2wj-Lj3DvrcNJQR7fttHIp0UUx3upHepsAJJTGgoTj5lzTBpONVKR8Ig97UrgdIKpaRfwdcHPUZ5JuY2qiRAno637jPraRWfqmAs4Tys4yGGTDyjIUx61GUU3skbRFUyAanv1tp-TZhFNOYAF0cFG4x4OoDz22LZGWVfXMlhpFH5u9eK4Vc0gM4DR-7aNfm7OAkfaw46nWuJY0sHPWQNnDC2QrgYj4QMeWj3IcvpvoXpNQ8Yg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Pulse-chase experiments of wild-type and mutant FV protein in COS-1 cells.
COS-1 cells, transiently transfected with pMT2/FV (wild type) or pMT2/FV-Arg2074Cys (mutant), were pulse-labeled with [35S]-methionine and [35S]-cysteine for 2 hours, and then chased by cold methionine and cysteine for various periods of time up to 180 minutes. At the specified chase period (0, 30, 60, 120, and 180 minutes) radiolabeled FV was immunoprecipitated from cell lysates (A,B) and from the corresponding conditioned media (C,D), electrophoresed on 4% SDS-PAGE gels under nonreducing conditions, and then detected by a phosphor imager (Typhoon 9200). The arrowheads indicate the 330-kDa FV molecule. In all panels, the pUC18 lane contains immunoprecipitable proteins at the end of the pulse period from COS-1 cells transfected with the unrelated pUC18 plasmid. Below each lane, a densitometric analysis of the band corresponding to immunoprecipitated FV is shown. Bar graphs are expressed as arbitrary densitometry units (y-axis), as calculated by the ImageQuant software by integrating intensities of all the pixels in the band excluding the background.
