Extended Data Fig. 5: Functional validations of iperoxo and QNB towards the WT or DREADD receptors.
From: Molecular basis for selective activation of DREADD-based chemogenetics
a—d, Binding affinities of iperoxo towards the WT and DREADD receptors. See Supplementary Table 5 for fitted parameter values that represent the mean ± SEM of n = 3 biologically independent experiments. e-f, Iperoxo, ACh, and DCZ agonist activities toward DREADD receptors by BRET2 assay. See Supplementary Table 6 for fitted parameter values that represent the mean ± SEM of n = 4 biologically independent experiments. g-h, Iperoxo, ACh, and DCZ agonist activities toward DREADD receptors by calcium flux assay (g) and cAMP Glo-sensor assay (h). See Supplementary Table 7 for fitted parameter values that represent the mean ± SEM of n = 3–5 biologically independent experiments. i-l, Gq activities of hM3R (i) and hM3Dq (j) under the stimulation of compounds ACh, DCZ, QNB, TIO, and 384 by FLIPR calcium assay and Gi/o activities of hM4R (k) and hM4Di (l) under the stimulation of compounds ACh, DCZ, QNB, tiotropium (TIO), AD-DX 384 (384) by split-luciferase-based cAMP Glo-sensor assay. See Supplementary Table 8 for fitted parameter values that represent mean ± SEM of n = 3 biologically independent experiments. m-p, Mutagenesis analysis of three arginines (R177, R184, and R552) in the ACh-induced activation for hM3R (m-n) and DCZ-induced activation for hM3Dq (o-p) by the BRET2 assay, respectively. See Supplementary Table 9 for fitted parameter values that represent mean ± SEM of n = 4 biologically independent experiments.
