Fig. 5
NPY displacement revealed biphasic characteristics and overexpression of Gαi shifted receptors towards the high-affinity state. a125I-PYY binding studies in transiently transfected Y2R HEK293 cells after incubation with 1 μM NPY, acidic wash and receptor recovery revealed diminished binding capacity of recycled receptors (n ≥ 3). b Specific 125I-PYY ligand displacement experiments with transiently transfected Y2R HEK293 cells showed a biphasic binding mode with a high affinity state (black line), which is lost after the first stimulation with 1 μM NPY, followed by acidic wash and recovery in ligand free medium for 60 min (olive-green line). For normalizing the data, total binding and unspecific binding of each condition were constrained to 100 and 0% respectively. Overexpression of Gαi shifted the percentage of the receptor towards the active state, which is summarized and represented in c). Experiments represent data n ≥ 3