Figure 1. Signaling pathways downstream of AT₁ receptor activation.

A. Various downstream molecules involved in the AT₁ receptor signaling, PLC: Phospholipase C, MAPKK: Mitogen activated protein kinase kinase, IP3: Inositol (1,4,5) P3, DAG: Diacyl glycerol, NFAT: Nuclear Factor of Activated T cells, EgR: immediate early growth response factor. Reporter constructs used to detect AT₁ receptor activation. B. Construction of a cell line that reports the activation of AT₁ receptors with increased luciferase activity Ang II regulation of 2XcEgR and 4XNFAT luciferase reporter constructs in AT₁ receptor expressing CHO cells. CHO.AT₁A cells were plated in either 12-wells or 24-wells plates and transiently transfected with 2XcEgR and 4XNFAT reporter constructs along with synthetic Renilla luciferase reporter (internal control) using Fugene 6 transfection reagent (Roche diagnostics, IN) for 6 hrs. Cells were serum starved for 24 hrs, treated with different concentrations of Ang II O/N, harvested and measured for relative luciferase activityusing a luminometer. Graph points denote the Relative Light Units (RLU).