Figure 2. Apelin 13-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/ AGTRL1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to stimulation with AGTRL1 receptor agonist, apelin 13. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (∆RFU) were normalized and plotted against the log of the cumulative doses (5-fold dilution) of apelin 13 (Mean ± SEM, n = 3). The EC50 of apelin 13 on this cell was 0.18 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 with a sigmoid shape.

Figure 1. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with Apelin-13 in CHO-K1/AGTRL1/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/AGTRL1/Gα15 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 Apelin-13 on CHO-K1/AGTRL1/Gα15 cells was 9.11 nM.
CHO-K1/AGTRL1/Gα15 Stable Cell Line
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