Figure 7.

Dose-response curves of ERK1/2 phosphorylation and cAMP production upon CCR9 and DRD5 stimulation. Jurkat cells were transfected with CCR9 and DRD5 and incubated for 48 hours. (A, B) Afterward, cells were preincubated in serum-free medium for 2 hours, seeded in white ProxiPlate 384-well microplates (3 × 10⁴ cells/well) and then treated with increasing concentrations of CCR9 agonist (CCL25) or DRD5 agonist (SKF81297) for 7 minutes. ERK1/2 phosphorylation was then determined by alpha-screen bead-based technology. Data are represented as % of phosphorylation of total ERK1/2. (C, D) Cells were preincubated in serum-free medium for 4 hours, seeded in white ProxiPlate 384-well microplates (10³ cells/well) and then treated with increasing concentrations of CCR9 agonist (CCL25) or DRD5 agonist (SKF81297) for 15 minutes. cAMP production was quantified by a TR-FRET methodology. Data are represented as % of cAMP accumulation. Each symbol represents data obtained from an individual determination (A, B: n = 6 determinations per group; C, D: n = 4–8 determinations per group). Mean ± SD are indicated. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001 (vs basal) by 1-way ANOVA followed by Tukey’s post hoc test.