For in vivo studies, lineage-negative (lin) cells were isolated from the bone marrow of 8- to 10-week-old 129SvEv mice, and murine PML-RAR cells were generated using a retroviral transduction and transplantation approach as previously described (33). The animals were checked periodically for clinical signs of disease and for presence of blast cells by May-Grünwald-Giemsa staining of blood smears. All-trans retinoic acid (ATRA) pellets (21-day release, 5 mg) or placebo pellets were implanted subcutaneously by trocar injection 10 days after cell injection. DDP_38003 was dissolved in a vehicle composed of 40% polyethylene glycol, molecular weight 400 in a 5% glucose solution and orally administered at the dose of 17 mg/kg 2 days per week, for 3 weeks. For the combination, mice were dosed with ATRA pellets (21-day release, 5 mg) and DDP_38003 at the dose of 17 mg/kg following the administration schedule of the single treatment. In vivo studies were performed after approval from our fully authorized animal facility and our institutional welfare committee Organismo Preposto al Benessere degli Animali (OPBA) and notification of the experiments to the Ministry of Health (as required by the Italian law; Institutional Animal Care and Use Committee numbers: 759/2015 in accordance with European Union directive 2010/63).

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