
Figure 1. Angiotensin II-induced concentration-dependent stimulation of intracellular calcium mobilization in HEK293/AT1 cells. The cells were loaded with Calcium-4 (Cat. No. R8141; Molecular Devices) prior to being stimulated with agonist angiotensin II. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of angiotensin II (Mean ± SEM, n = 3). The EC50 of angiotensin II on this cell was 1.23 nM.
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 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with angiotensin II in HEK293/AT1 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in HEK293/AT1 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of angiotension II on HEK293/AT1 cells was 3.53 nM.
HEK293/AT1 Stable Cell Line
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