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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