
Figure 1. CXCL12-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/CXCR4 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with CXCR4 agonist, CXCL12 (Cat. No. Z02825, Genscript). The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of CXCL12 (mean ± SEM, n = 3). The EC50 of CXCL12 on CHO-K1/Gα15/CXCR4 cells was 0.40 μg/ml.
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 CXCL12 in CHO-K1/Gα15/CXCR4 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/CXCR4 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 CXCL12 on CHO-K1/Gα15/CXCR4 cells was 20.76 ng/ml.
CHO-K1/CXCR4/Gα15 Stable Cell Line
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