Table 2.
Summary of pEC50 values and relative efficacy of NECA and HEMADO at A3-YFP and A3 W243F-YFP in second-messenger assays
| pERK1/2 assay | cAMP accumulation assay | CRE-SPAP assay | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| pEC50 | Maximum response (% 10 μM NECA) | n | pEC50 | Maximum response (% inhibition of 3 μM FSK) | n | pEC50 | Maximum response (% inhibition of 10 μM FSK) | n | ||
| NECA | A3-YFP | 7.77 ± 0.09 | 100 | 4 | 7.57 ± 0.15 | 77.5 ± 6.9 | 4 | 7.65 ± 0.23 | 65.8 ± 8.2 | 4 |
| A3 W243F-YFP | 7.65 ± 0.11 | 22.4 ± 3.3 | 4 | 7.53 ± 0.16 | 49.7 ± 2.0 | 4 | 7.44 ± 0.24 | 34.8 ± 6.7 | 5 | |
| HEMADO | A3-YFP | 8.72 ± 0.02 | 119.5 ± 5.8 | 4 | 8.62 ± 0.12 | 80.0 ± 5.1 | 4 | 8.52 ± 0.6 | 73.2 ± 7.7 | 9 |
| A3 W243F-YFP | 7.71 ± 0.21 | 16.0 ± 3.1 | 4 | 8.02 ± 0.22 | 43.2 ± 4.3 | 4 | NR | No inhibition | 9 | |
Values are mean ± SEM from n separate experiments. Maximum response in pERK1/2 assay is the response in each cell line relative to that of 10 μM NECA in A3-YFP-expressing cells. In CRE-SPAP and [3H]-cAMP assays, maximal response is the maximal inhibition of forskolin-stimulated CRE-SPAP expression or accumulation achieved with each agonist in each cell line. [3H]-cAMP data represents measurements taken after 1 h stimulation. NR, no response.