Hy y1888

1000x stocks of ABA (100 mM, Gold Biotechnology A-050-500) and GA (10 mM, Toronto Research Chemicals G377500) were prepared by dissolving in 100% ethanol and stored at −20 °C. 10000x stocks of Dano (10 mM, MedChem Express HW-10238) and CHIR 90021 (GSK3 inhibitor) (10 mM, Tocris Bio-Techne 4423) were prepared by dissolving in dimethyl sulfoxide (DMSO) and stored at −20 °C. 50000x stocks of 4OHT (50 mM HelloBio HB2508) were prepared by dissolving in DMSO and stored at −20 °C. For in vitro induction, the stock solutions were diluted to 5X with culture media and then added to the cell cultures such that the final induction concentration is 1X. For in vivo administration, GA was dissolved to 2 mg/ml in 2.5% DMSO/PBS and ABA was dissolved to 20 mg/ml in 2.5% DMSO/corn oil (MedChemExpress HY-Y1888. Mice received 100ul inducer for each induction.

Thirty-six adult male C57BL/6 mice were divided into six groups according to a computer-based randomization in experiment 1 (n = 12) as follows: 1) sham + vehicle, 2) ICH + vehicle, and 3) ICH + Dex. Autologous blood injection in the basal ganglia was used to establish a mild ICH model in mice. Based on previous studies (Gu et al., 2011 (link); Liang et al., 2017 (link)) and our preliminary experiments, 25 μg/kg of Dex (Jiangsu Hengrui Medicine Co., Ltd., Jiangsu, China) was administered using intraperitoneal injection post-ICH daily for 3 days. Mice in sham + vehicle and ICH + vehicle groups received an equal volume of saline via intraperitoneal injection (Figure 1A).
Twenty-four adult male C57BL/6 mice were divided into two groups according to a computer-based randomization in experiment 2 (n = 12) as follows: 1) ICH + Dex + GSK and 2) ICH + Dex + vehicle. GSK1016790A (GSK, 1 μmol/2 μl, HY-19608, MedChemExpress, United States), an agonist of TRPV4, was dissolved in 10% dimethyl sulfoxide (DMSO) (D8418, Beyotime, China) and 90% corn oil (HY-Y1888, MedChemExpress, United States), and injected into the left lateral ventricle (stereotaxic coordinates: 1.0 mm lateral to the midline, 0.3 mm posterior to the bregma, and 2.5 mm below the skull) 30 min before Dex administration. Vehicle was administered with an equivalent volume of DMSO plus corn oil as a control (Figure 1B).