Figure 1. Oxotremorine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/M2 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M2 receptor agonist, Oxotremorine. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (10-fold dilution) of Oxotremorine (Mean ± SD, n = 2). The EC50 of Oxotremorine on this cell was 63.7 nM.
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.
Figure 2. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of [3H]N-Methylscopolamine ([3H]NMS) in the absence (total binding) or presence of 1000- fold access unlabeled Atropine (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. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of Atropine in the presence of 0.2 nM [3H]N-Methylscopolamine ([3H]NMS). Binding was terminated by rapid filtration. Data were fit to one-site competition equation using a non-linear regression method.
CHO-K1/M2/Gα15 Stable Cell Line
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