Figure 4
Figure 4. Effect of warfarin on the secretion of PZ and FX in HEK 293 cells. (A) HEK 293 cells transiently transfected with a PZ or FX cDNA were treated with or without 10 μg/mL warfarin and/or 5 μg/mL Vit.K for 24 hours. γ-Carboxylated PZ or FX in 0.2 mL medium was collected by barium citrate adsorption in a 30-μL solution. Medium or barium citrate adsorbate (10 μL) was subjected to SDS-PAGE/Western blot analysis. (B) HEK 293 cells transfected with PZMetLuc (left, circles) or FXMetLuc (right, squares) were divided into 6 wells on a 48-well plate at 48 hours after transfection and treated with 0, 0.01, 0.1, 1, 10, or 100 μg/mL warfarin in the presence of 5 μg/mL Vit.K for 24 hours. Luciferase activity in the medium (open symbols) and barium citrate-adsorbate (closed symbols) were calculated relative to the activity of the samples without warfarin treatment (arbitrarily set at 100%).

Effect of warfarin on the secretion of PZ and FX in HEK 293 cells. (A) HEK 293 cells transiently transfected with a PZ or FX cDNA were treated with or without 10 μg/mL warfarin and/or 5 μg/mL Vit.K for 24 hours. γ-Carboxylated PZ or FX in 0.2 mL medium was collected by barium citrate adsorption in a 30-μL solution. Medium or barium citrate adsorbate (10 μL) was subjected to SDS-PAGE/Western blot analysis. (B) HEK 293 cells transfected with PZMetLuc (left, circles) or FXMetLuc (right, squares) were divided into 6 wells on a 48-well plate at 48 hours after transfection and treated with 0, 0.01, 0.1, 1, 10, or 100 μg/mL warfarin in the presence of 5 μg/mL Vit.K for 24 hours. Luciferase activity in the medium (open symbols) and barium citrate-adsorbate (closed symbols) were calculated relative to the activity of the samples without warfarin treatment (arbitrarily set at 100%).

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