
Figure 1. SNC80-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRD1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with OPRD1 agonist, SNC80. 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 SNC80 (mean ± SEM, n = 3). The EC50 of SNC80 on CHO-K1/OPRD1/Gα15 cells was 0.75 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 SNC80 in CHO-K1/OPRD1/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/OPRD1/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 SNC80 on CHO-K1/OPRD1/Gα15 cells was 0.10 μM.
CHO-K1/OPRD1/Gα15 Stable Cell Line
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