
Figure 1. Calcitonin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/AMY1 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with AMY1 agonist, calcitonin. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of calcitonin (mean ± SEM, n = 3). The EC50 of calcitonin on CHO-K1/Gα15/AMY1 cells was 0.11 μM.
Notes:
EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom) / (1+10^ ((LogEC50-X)*Hill Slope))
X is the logarithm of concentration. Y is the response.
Y is RFU and starts at Bottom and goes to Top along a sigmoid curve.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with amylin in CHO-K1/Gα15/AMY1 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/AMY1 cells competitively bind with Europium Cryptate-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of amylin on CHO-K1/Gα15/AMY1 cells was 4.94 nM.
CHO-K1/Gα15/AMY1 Stable Cell Line
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