Effect of Various Inhibitors of Protein Degradation and Brefeldin A on Intracellular Levels of FVIIwt, FVIICys97, and FVIIArg100
| . | FVIIwt . | FVIICys97 . | FVIIArg100 . |
|---|---|---|---|
| Media alone (n = 6) | 100 ± 1.1 | 100 ± 3.1 | 100 ± 4.2 |
| NH4Cl, 50 mmol/L (n = 3) | 107 ± 4.2 | 83.4 ± 3.6 | 113 ± 2.7 |
| Leupeptin, 100 μmol/L (n = 3) | 98 ± 1.2 | 100.5 ± 3.9 | 84 ± 3.6 |
| Lactacystin, 10 μmol/L (n = 3) | 88 ± 1.6 | 91.4 ± 2.8 | 98.1 ± 5.5 |
| ALNN, 50 μg/mL (n = 9) | 96.4 ± 5.7 | 147 ± 8.93-150 | 113 ± 4.1 |
| Brefeldin A, 10 μg/mL (n = 6) | 190 ± 10.43-151 | 125 ± 7.53-152 | 151 ± 10.83-153 |
| . | FVIIwt . | FVIICys97 . | FVIIArg100 . |
|---|---|---|---|
| Media alone (n = 6) | 100 ± 1.1 | 100 ± 3.1 | 100 ± 4.2 |
| NH4Cl, 50 mmol/L (n = 3) | 107 ± 4.2 | 83.4 ± 3.6 | 113 ± 2.7 |
| Leupeptin, 100 μmol/L (n = 3) | 98 ± 1.2 | 100.5 ± 3.9 | 84 ± 3.6 |
| Lactacystin, 10 μmol/L (n = 3) | 88 ± 1.6 | 91.4 ± 2.8 | 98.1 ± 5.5 |
| ALNN, 50 μg/mL (n = 9) | 96.4 ± 5.7 | 147 ± 8.93-150 | 113 ± 4.1 |
| Brefeldin A, 10 μg/mL (n = 6) | 190 ± 10.43-151 | 125 ± 7.53-152 | 151 ± 10.83-153 |
90% confluent stably transfected CHO cells expressing recombinant FVIIwt, FVIICys97, or FVIIArg100 were incubated 4 hours in fresh media containing 10% FBS in the presence or absence of the different agents. The figures in parentheses are the number of independent transfections performed on cells from a single clone expressing each construct. The levels of VII:Ag were then measured in cell lysates and the results are expressed as the percentage of FVII (mean ± SE) produced in the absence of the agent.
P = .0004.
P = .0001.
P = .02.
P = .002.