Daunorubicin dnr

Chidamide (CS055, purity >95%) was supplied by Chipscreen Bioscience Ltd. (Shenzhen, China) and dissolved in dimethyl sulfoxide (DMSO) (Invitrogen, USA) to obtain a stock solution of 50 mM. Idarubicin (IDA), daunorubicin (DNR), and cytarabine (Ara-C) were purchased from Sigma-Aldrich (St Louis, MO) and dissolved in phosphate-buffered saline (PBS) at the concentrations of 10, 100, and 100 mM, respectively. All the above stock solutions were stored at −20 °C. It was diluted to the required concentrations in subsequent experiments with culture medium.

The pan-Pim kinase inhibitor AZD1208, from Tocris Bioscience (Minneapolis, MN, USA), was used at 1 μM based on phase I clinical trial data [10 (link)] and on inhibition of BAD phosphorylation at serine 112 as a pharmacodynamic endpoint [37 (link)]. The topoisomerase 2 inhibitors daunorubicin (DNR), mitoxantrone (MXR) and etoposide (VP-16) (Sigma-Aldrich) were dissolved in dimethylsulfoxide (DMSO) (less than 0.05 percent) and used at their approximate IC₅₀ concentrations of 10, 20 and 30 nM, respectively, in FLT3-ITD cell lines [13 (link)]. The protein translation inhibitor cycloheximide (CHX) was from Sigma-Aldrich and the proteasome inhibitor carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG-132) from Calbiochem (San Diego, CA, USA). The c-Myc inhibitor 5-[(4-ethylphenyl)methylene]-2-thioxo-4-thiazolidinone (10058-F4), which inhibits c-Myc-Max binding and thereby prevents transcriptional activation of c-Myc targets, was also from Sigma-Aldrich.