C57Bl/6 and MX1-Cre mice were obtained from The Jackson Laboratory. Dyrk1a C57Bl/6 mutant mice have been described previously (17). Genotyping of Dyrk1a floxed or WT alleles was performed using PCR primers (forward, 5′-ATTACCTGGAGAAGAGGGCAAG-3′ and reverse, 5′-TTCTTATGACTGGAATCGTCCC-3′). For xenograft experiments, NOD.Cg-PrkdcscidIl2rgtm1Wjl SzJ (NSG) mice were purchased from The Jackson Laboratory. Mice were maintained in specific pathogen–free conditions. Littermate controls were used for analysis in each cohort. Animals were backcrossed at least 10 generations. Male and female mice between birth and 1 year of age were studied.

Animal studies were performed to determine the genetic requirement of Dyrk1a in murine models of B-ALL, the on-target effects of EHT 1610 and AS1842856, and the efficacy of EHT 1610 and AS1842856 in treating mouse models of B-ALL. The sample size of the animal experiments, the specific transplantation protocols, and the dosing are described in the figure legends and in the appropriate Methods sections, respectively. For all in vivo experiments, mice were randomly assigned to transplant groups (genetic models) or treatment groups (inhibitor studies). All experiments were performed in an unblinded manner.

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