Embryos were dechorionated and resuspended in 1 mL of 1X E3 media with 0.004% 1-phenyl-2-thiourea (PTU) and incubated with Venetoclax or Flavopiridol (MedChem Express) at the desired drug concentration in 12-well dishes beginning at 24 hpf. Dimethyl sulfoxide (DMSO) was used to solubilize the drugs and was also used as a vehicle for control experiments. Drug treatments were continuous at 28˚C until 72 hpf when embryos were fixed in 4% PFA/PBS.
For recovery experiments, lyz:MLL-ENL injected embryos were dechorionated and split evenly in a 12 well dish. Embryos were then incubated in 1X E3 media and 0.004% PTU containing either 200 nM Venetoclax, 200 nM Flavopiridol, 200 nM Venetoclax and Flavopiridol, or DMSO beginning at 24 hpf at 28˚C before being fixed in 4% PFA/PBS at 72 hpf.
For recovery experiments, lyz:MLL-ENL injected embryos were dechorionated and split evenly in a 12 well dish. Embryos were then incubated in 1X E3 media and 0.004% PTU containing either 200 nM Venetoclax, 200 nM Flavopiridol, 200 nM Venetoclax and Flavopiridol, or DMSO beginning at 24 hpf at 28˚C before being fixed in 4% PFA/PBS at 72 hpf.
