Cotransfections with wild-type and mutant fXI expression constructs. The 293 cells were cotransfected with 0.08 μg wild-type fXI construct and 0.08 μg pJVCMV (lane 1), 0.08 μg additional wild-type fXI construct (lane 2), or 0.08 μg mutant fXI constructs (lanes 3-7). Cells transfected with 0.16 μg pJVCMV without cDNA insert (lane 8) were the negative control. Shown are relative concentrations of protein in media (A) and cell lysates (B) 67 hours after transfection, compared with the means for wild-type fXI (lane 1), which were assigned a value of 100%. Results are means and standard deviations for 9 separate transfections, each tested in triplicate by ELISA. Results are corrected for transfection efficiency using a Renilla luciferase assay. Lanes are as follows: (1) Wild-type (WT)–fXI + pJVCMV; (2) WT-fXI + WT-fXI; (3) WT-fXI + fXI-Glu117Stop; (4) WT-fXI + fXI-Phe283Leu; (5) WT-fXI + fXI-Gly350Glu; (6) WT-fXI + fXI-Gly400Val; (7) WT-fXI + fXI-Trp569Ser; and (8) pJVCMV control.