Effects of various inhibitors on the secretion and degradation of wild-type and Tenri-type factor XIIs. Stably transfected BHK cells expressing wild-type (left panel) or Tenri-type (right panel) factor XII were pulse-labeled for 30 minutes with 100 μCi/mL EXPRE35S35S and then chased for 16 hours in the presence of various inhibitors. Factor XII in the cell extracts and culture media were subjected to immunoprecipitation followed by SDS-PAGE analysis. Lane 1, sample from pulse-labeled cells; lane 2, sample from 16-hour chased cells without inhibitors; lanes 3 through 9, samples from 16-hour chased cells in the presence of inhibitors; lane 3, 5 μg/mL brefeldin A; lane 4, 100 μmol/L chloroquine; lane 5, 100 μg/mL leupeptin; lane 6, 100 μmol/L E64d; lane 7, 20 μmol/L LL; lane 8, 20 μmol/L LLL; and lane 9, 20 μmol/L lactacystin. Positions of factor XII are indicated by arrows.
