Experimental design for kinetic dissociation experiments
Binding of [125I]RTI-55 (0.01 nM) to hDAT proceeded for 2 h at 25°C. At this point (time 0), RTI-55 (1 μ M) was added to conditions 2 and 4. Ten minutes later, test drugs were added to conditions 3 and 4. For the data analysis, the 100% of control point was time 0 min for conditions 1 and 2 and time 10 min for conditions 3 and 4. Each data point is the mean ± S.D. of three experiments. The data were fit to a one- (conditions 1, 2, and 4) or two-component (condition 3) exponential decay model using KaleidaGraph version 3.6 to calculate the kinetic constants.
Condition | First Addition (Time 0 min) | Second Addition (Time 10 min) |
|---|---|---|
| 1. Control | None | None |
| 2. RTI-55 | RTI-55 (1 μm) | None |
| 3. Test drug | None | Test drug: SoRI-20040 (10 μM) or SoRI-20041 (10 μM) or SoRI-2827 (10 μM) |
| 4. RTI-55 + test drug | RTI-55 (1 μM) | Test drug: SoRI-20040 (10 μM) or SoRI-20041 (10 μM) or SoRI-2827 (10 μM) |