Hb2508

MTZ (MilliporeSigma, M3761-25G) was dissolved in system water at a concentration of 5 mM and 0.5% DMSO. Adult zebrafish were exposed to MTZ for 16 hours at ambient temperature in the dark. APAP (MilliporeSigma, A7085-100G) was dissolved in system water at a concentration of 15 mM. EtOH (MilliporeSigma, EX0276-3) was dissolved in system water at a concentration of 1%. Adult zebrafish were exposed to APAP or EtOH for 24 hours at ambient temperature in the dark. Stock solutions for AG1478 (Selleck Chemicals, S2728), U0126 (Cayman Chemical Company, 70970), LY290002 (Selleck Chemicals, S1105), and Rapamycin (Selleck Chemicals, AY-22989) were made by dissolution in DMSO at a concentration of 10 μM, 20 μM, 10 μM, and 10 μM, respectively. These chemicals were diluted 1:1,000 in system water to make a working solution. Adult zebrafish were immersed in the working solution. Working solution was changed daily. 4-OHT (Hello Bio, HB2508) was dissolved in DMSO at a concentration of 10 mM, vortexed continuously for 15 minutes, and stored at –20°C. Prior to use, an aliquot of 4-OHT was incubated at 65°C for 10 minutes. Triple-transgenic larvae were exposed to 4-OHT between 2 and 3 days after fertilization to induce recombination.

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.