Fig. 2.
Fig. 2. Secretion and thrombin cleavage of WT and mutant FVIII. The secretion of each mutant was analyzed by immunoprecipitation of conditioned medium from [35S]-methionine pulse-labeled transfected cells chased for 4 hours in medium containing excess unlabeled methionine. Immunoprecipitated FVIII molecules were analyzed by SDS-PAGE before (−) and after (+) thrombin (IIa) digestion. Mock indicates cells that did not receive plasmid DNA. Molecular weight size markers are shown on the left. Single chain (Single), heavy chain (Heavy), and light chain (Light) are indicated for undigested samples. A3-C1-C2, A1, and A2 fragments are indicated for digested samples. The symbol * represents the A2 fragment with reduced mobility.

Secretion and thrombin cleavage of WT and mutant FVIII. The secretion of each mutant was analyzed by immunoprecipitation of conditioned medium from [35S]-methionine pulse-labeled transfected cells chased for 4 hours in medium containing excess unlabeled methionine. Immunoprecipitated FVIII molecules were analyzed by SDS-PAGE before (−) and after (+) thrombin (IIa) digestion. Mock indicates cells that did not receive plasmid DNA. Molecular weight size markers are shown on the left. Single chain (Single), heavy chain (Heavy), and light chain (Light) are indicated for undigested samples. A3-C1-C2, A1, and A2 fragments are indicated for digested samples. The symbol * represents the A2 fragment with reduced mobility.

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