Fig. 3: Profiling of µ-opioid receptor (MOP) agonizts and antagonists using the 7TM phosphorylation assay. | Communications Biology

Fig. 3: Profiling of µ-opioid receptor (MOP) agonizts and antagonists using the 7TM phosphorylation assay.

From: A bead-based GPCR phosphorylation immunoassay for high-throughput ligand profiling and GRK inhibitor screening

Fig. 3

a Schematic representation showing the MOP binding sites for antibodies against phosphorylated (p)T370-, pS375-, pT376-, and pT379-MOP and MOP independent of phosphorylation (np-MOP). b–f Concentration–response curves were generated after treatment with DAMGO (b), naloxone (c), morphine (d), fentanyl (e) and the PKC activator phorbol 12-myristate 13-acetate (PMA) (f). The cells were exposed to agonizts or PMA for 30 min at 37 °C. The antagonist naloxone was added 30 min prior to DAMGO stimulation. The cells were then lysed and analyzed according to the standard assay protocol. Every graph represents the mean of n = 5 independent experiments performed in duplicates ± SEM. Where error bars are not apparent, SEM was smaller than symbol size. All data points were normalized to 10 µM DAMGO stimulation.

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