Experimental animals and models
Male FVB wild-type (WT) mice aged between 8-12 weeks were randomized, grouped, and used for both in vitro and in vivo experiments in this study. PCH in vivo model was made by subcutaneous injection of Isoproterenol (160504; Sigma) 0.5mg/100g/day for 40 days. An equivalent volume of the placebo (5% v/v dimethyl sulfoxide (DMSO)) was injected subcutaneously to the vehicle (Vhl) group. Their littermates without any handling were included in the study and designated as the control (Ctrl) group.
PCH preventive models were made by intraperitoneal injection of a GRK5 inhibitor drug, Amlexanox (ab142825; Abcam) 2.5mg/100g/day or, and ACs direct activator Forskolin (1099; Tocris Bioscience, UK) 0.5mg/100g/day, simultaneously with ISO 0.5mg/100g/day from day 1 to day 40 (Supplementary Fig. S1).