Fig 1: In vitro potency of sartans at blocking AT1R-induced G protein or βarr activation.(A) Dose-response curves for inhibition of AngII-induced AT1R-βarr binding or for inhibition of AngII-induced AT1R-G protein coupling (early response, ER) for each compound, as derived from the CellKey assay. (B) Dose-response curves of each drug for βarr activation at the AngII-bound AT1R, based on the DiscoveRx PathHunterTM β-Arrestin assay system. (C) AngII-induced G protein inhibition potencies based on the FLIPR calcium assay (Molecular Devices). Los: Losartan; Exp: EXP3174; Tel: Telmisartan; Epr: Eprosartan; Azi: Azilsartan; Olm: Olmesartan; Can: Candesartan; Val: Valsartan; Irb: Irbesartan.
Fig 2: L-162,313 is an AT1R partial agonist. (A) Chemical structures of small-molecule ligands for AT1R. (B) Activation of G⍺q mediated inositol monophosphate in response to L-162,313 and AngII (IPone, CisBio). (C) G⍺q mediated calcium mobilization (FLIPR Calcium 6, Molecular Devices). (D) AT1R mediated dissociation of the Gq heterotrimer monitored via BRET (9). (E) L-162,313 and AngII desensitize calcium mobilization upon rechallenge with AngII. (F) Recruitment of Nluc-β-arrestin2 to AT1R-mVenus. (G) Endosomal translocation of Nluc-β-arrestin2 to FYVE-linked mVenus. Error bars represent the mean SE from at least three independent experiments. EV represents empty vector control to ensure signals are receptor dependent.
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