Table 3.
Critical Graphs From Competition Assays in U2OS Cells With GR Induction of GREtkLUC Reporter
| Chemical (F2) | 1/EC50 vs |
Amax/EC50 vs |
EC50/Amax vs |
Mechanism | |||
|---|---|---|---|---|---|---|---|
| F1 | F2 | F1 | F2 | F1 | F2 | ||
| AC93253 | Slope = 0 (−0.000046 ± 0.010, SD, n = 20, 5 experiments), y-axis intercept < with F2 | Lines intersect at origin, slope < with F2 | Linear, slope < with F1 | GREtkLUC is A = CLS AC93253 is C ≤ CLS | |||
| Baclofen | Slope = 0 (−0.00075 ± 0.0105, SD, n = 12, 3 experiments) | Lines intersect at origin, slope ≤ with F2 | Linear, slope < with F1 | GREtkLUC is A = CLS Baclofen is a weak C ≤ CLS | |||
| Camptothecin | Slope = 0 (−0.0044 ± 0.0070, SD, n = 12, 3 experiments) | Lines intersect at origin, slope < with F2 | Linear, slope < with F1 | GREtkLUC is A = CLS Camptothecin is C ≤ CLS | |||
| Dihydroouabain | slope = 0 (−0.0017 ± 0.016, SD, n = 24, 6 experiments), y-axis intercept < with F2 | Lines intersect at origin, slope > with F2 | Lines intersect at F2 < 0, slope > with F1 | GREtkLUC is A = CLS Dihydroouabain is A > CLS | |||
| Emetine | Slope = 0 (−0.0012 ± 0.0074, SD, n = 16, 4 experiments) | Lines intersect at origin, slope < with F2 | Plots curve up, position < with F1 | GREtkLUC is A = CLS Emetine is C at 2 sites ≤ CLS | |||
| Nocodazole | Slope = 0 (0.0054 ± 0.020, SD, n = 12, 3 experiments), y-axis intercept > with F2 | Lines intersect at origin, slope > with F2 | Lines intersect at F2 < 0, slope > with F1 | GREtkLUC is A = CLS Nocodazole is A > CLS | |||
| NU6027 | Slope = 0 (−0.0023 ± 0.015, SD, n = 12, 3 experiments) | Lines intersect at origin, slope > with F2 | Lines intersect at F2 < 0, slope > with F1 | GREtkLUC is A = CLS NU6027 is A > CLS, | |||
| Phenanthroline | Slope = 0 (−0.011 ± 0.026, SD, n = 20, 5 experiments) | Lines intersect at origin, slope > with F2 | Lines intersect at F2 < 0, slope > with F1 | GREtkLUC is A = CLS Phenanthroline is A > CLS | |||
| Sanguinarine | slope = 0 (0.00026 ± 0.0060, SD, n = 34, 9 experiments), y-axis intercept < with F2 | Lines intersect at origin, slope < with F2 | Linear, slope < with F1 | GREtkLUC is A = CLS Sanguinarine is C ≤ CLS | |||
| Stattic | Slope = 0 (0.0021 ± 0.000196, SD, n = 16, 4 experiments), y-axis intercept < with F2 | Lines intersect at origin, slope < with F2 | Linear, slope < with F1 | GREtkLUC is A = CLS Stattic is C ≤ CLS | |||
A, accelerator; C, competitive decelerator; CLS, concentration limiting step (see text for explanations). GREtkLUC = factor F1 in all entries. The 3 types of graphs (1/EC50, Amax/EC50, and EC50/Amax) vs each factor are listed at the top, with the characteristics of the most informative graphs vs F1 (GREtkLUC) or vs F2 (chemical), listed below the relevant factor. In these columns, “>” and “<” mean “increases” and “decreases” respectively. The unique mechanistic conclusion for each pair is listed at the far right under “Mechanism.” In this column, “ =,” “<,” and “≤” mean “at,” “before,” and “before or at,” respectively.