2.10 Animal model studies
All animal care and experimental techniques for anesthetics, hair regrowth experiments and assays were accomplished with the permission of Animal Research Ethics Committee of Shenzhen University (permit number AEWC-201412003). Animal studies are stated in amenability with the ARRIVE guidelines and with the commendations made by the British Journal of Pharmacology (Lilley et al., 2020) Specific pathogen-free C57BL/6 mice (6 weeks old) were purchased from Nanjing Biomedical Research Institute of Nanjing University, (Nanjing, China) and acclimatized for 1 week. After anesthesia via an intraperitoneal injection of 2,2,2-tribromoethanol (Sigma-Aldrich, St. Louis, USA), the skin hairs on the dorsal side of 7-week-old mice (whose hair follicles were in the telogen phase) were clean-shaven by a hair clipper. The mice were randomly separated into three groups(Lee et al., 2012).. For every mouse, 200 μL of each drug was topically applied at an appropriate concentration everyday (as described in figure legends) for 25 days. The control group received vehicle alone, the ISX9 group received vehicle containing 1% of ISX9, and the minoxidil group received 2% of minoxidil. All reagents used for the hair growth test were dissolved in a vehicle composed of 50% (v/v) ethanol, 30% water, and 20% propylene glycol. The back skin of the mice was photographed at 0, 7, 14, 21, 25 days, and analyses was done regarding the weight of regrown hair in different groups treated for 25 days. After killing mice, skin tissues were gathered and saved at -80 °C.