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).