Figure 1. SNC80-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRD1/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist SNC80. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of SNC80 (Mean ± SD, n = 2). The EC50 of SNC80 on this cell was 0.75 µM.
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
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.
2. Signal to background Ratio (S/B) = Top/Bottom
Figure 2. 5 μg of membranes prepared from CHO-K1 cells stably expressing OPRD1 receptors were incubated with indicated concentrations of [3H]Naltrindole in the absence (total binding) or presence of 1000-fold excess unlabeled Naloxone (nonspecific binding, NSB). Binding was terminated by rapid filtration. Specific binding was defined by subtracting NSB from total binding. Data were fit to one-site binding equation using a non-linear regression method.
Figure 3. 5 μg of membranes prepared from CHO-K1 cells stably expressing OPRD1 receptors were incubated with indicated concentrations of Naloxone in the presence of 0.4 nM [3H]Naltrindole. Binding was terminated by rapid filtration. Data were fit to one-site competition equation using a non-linear regression method.
CHO-K1/OPRD1/Gα15 Stable Cell Line
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