Fig. 10: Cerebral vasospasm and behavioral disorders are efficiently targeted by UT-biased ligand/antagonists.
a–e Antagonistic effect of palosuran, GSK1562590 and ligand bias of urantide on the activation of G proteins and β-arrestins on UII-activated UT signaling pathways (3 independent experiments for each in triplicate each). a Using BRET2 biosensors, UII activation of Gi coupled family members Gαi1, Gαi2, Gαi3, and Gαz was assessed in HEK293 cells transiently expressing hUT in the presence of 10 µM of the indicated biased ligand/antagonists. b The activation of the Gq pathway was accessed using both a BRET2 biosensor and by determining the production levels of the second messenger IP1 in the presence of 10 µM of the indicated biased ligand/antagonists. c Ca2+ mobilization induced by graded concentrations (10−11 to 10−6 M) of UII on hUT-transfected HEK-293 cells pretreated with buffer or biased ligand/antagonists. d Using BRET2 biosensors, UII activation of Go coupled family members GoA and GoB was assessed in HEK293 cells transiently expressing hUT in the presence of 10 µM of the indicated biased ligand/antagonists. e Using BRET2 biosensors, the UII-induced recruitment of β-arrestin 1 and β-arrestin 2 was assessed in HEK293 cells transiently expressing hUT in the presence of 10 µM of the indicated biased ligand/antagonists (a–e, n = 3; values are expressed as the mean ± SEM). f Timeline of intracisternal injection of palosuran, GSK1562590 or urantide during SAH induction prior to CVS analysis and behavioral testing. g Graphs (right panel) representing changes in lumen area/wall thickness on MCA (n = 6/condition). Values are expressed as the mean ± SEM. ***P < 0.001 for comparisons with aCSF + sham condition. $$$P < 0.001 for comparisons with aCSF + SAH condition (two-sided one-way analysis of variance (ANOVA), Bonferroni’s correction). Exploration in OFT (10 min) and sensorimotor functions in BWT (n = 6–10/condition). Values are expressed as the mean ± SEM *P < 0.05; **P < 0.01; ***P < 0.001 for comparisons with aCSF + sham condition. $P < 0.05; $$P < 0.01; $$$P < 0.001 for comparisons with aCSF + SAH condition (two-sided one-way analysis of variance (ANOVA), Bonferroni’s correction). Source data are provided as a Source Data file. f created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en).
