Prolonged Calcitonin Receptor Signaling by Salmon, but Not Human Calcitonin, Reveals Ligand Bias (original) (raw)
Figure 7
Prolonged ligand dissociation kinetics on membranes preparations and live cells.
A) Total binding of 125I-sCT and 125I-hCT ligand bound to U2OS CALCR cells at different time points (0, 2, 4, 8, 24, 48 and 72 hours). B0 is total 125I-ligand bound to cells at t = 0 and B is 125I-ligand bound. B) Specific activity of 125I-sCT and 125I-hCT ligand released into the supernatant by cells in (B) at different time points (0, 2, 4, 8, 24, 48 and 72 hours). C) 125I-sCT and 125I-hCT ligand dissociation was investigated using isolated U2OS CALCR membrane preparations. The dissociation from the membranes was measured after 1, 2, 4, 8, 24, 48 and 72 hours with or without the presence of 20 μM GTPγS to determine G-protein dependency. B0 is total 125I-ligand bound to membranes at t = 0 and B is 125I-ligand bound.B0 is total 125I-ligand bound to cells at t = 0 and S is 125I-ligand in the medium. Live cell assays were conducted at 37°C and membrane assays were conducted at room temperature.