For in vitro Mll1 deletion, Cre induction was initiated in culture medium supplemented with 100 nM 4-OHT dissolved in ethanol for 6 h. Deletion efficiency for all alleles was determined by semi-quantitative genomic PCR using primers described (Chen et al., 2017 (link)), at 24 h after deletion was initiated.
MENIN inhibitors MI-3454 and SNDX-5613 were purchased from MedChemExpress (Monmouth Junction, NJ) and dissolved in DMSO. WDR5 inhibitor OICR-9429 was purchased from Selleck Chemicals (Houston, TX). Primary mouse leukemia cells generated previously (Nguyen et al., 2016 (link)) were plated at 0.5 to 1 × 105 cells/mL in IMDM supplemented with 15% heat inactivated fetal bovine serum, 1x penicillin/streptomycin, mouse cytokines (100 ng/mL SCF, 6 ng/mL IL-3, and 10 ng/mL IL-6), and treated with the inhibitors at the indicated concentrations or control DMSO. Cell concentrations were restored to the starting concentrations and inhibitors replenished after 72 h.
MENIN inhibitors MI-3454 and SNDX-5613 were purchased from MedChemExpress (Monmouth Junction, NJ) and dissolved in DMSO. WDR5 inhibitor OICR-9429 was purchased from Selleck Chemicals (Houston, TX). Primary mouse leukemia cells generated previously (Nguyen et al., 2016 (link)) were plated at 0.5 to 1 × 105 cells/mL in IMDM supplemented with 15% heat inactivated fetal bovine serum, 1x penicillin/streptomycin, mouse cytokines (100 ng/mL SCF, 6 ng/mL IL-3, and 10 ng/mL IL-6), and treated with the inhibitors at the indicated concentrations or control DMSO. Cell concentrations were restored to the starting concentrations and inhibitors replenished after 72 h.