Fig. 2

The Menin-MLL1 inhibitor induced differentiation and loss of LICs of human MLL-AF9–evoked murine AML-like cells A RT‒qPCR was performed in MLL-AF9–evoked murine AML-like cells after treatment with DS-1594a·HCl (1.5, 4.6, 14, and 41 nM) for 4 days. The expression levels of Meis1, Hoxa9, Mef2c, and Pbx3 were normalized to that of Actb and compared to those in the DMSO-treated control (mean ± SD; n = 3). B FCM analysis with the indicated antibodies (cKit, CD11b, Ly-6G, and Annexin V) for MLL-AF9–evoked murine AML-like cells after 7 days of treatment with DMSO, DS-1594a·succinate (10, 20, and 40 nM), or Ara-C (50 and 100 nM). The bars represent the mean ± SD; n = 3. C Serial replating in the MethoCult M3434 assay. The colonies were counted 4–5 days after seeding with DMSO, DS-1594a·HCl (10, 20, and 40 nM), or Ara-C (50 and 100 nM). The first colony (MC1) was counted, collected, and replated (MC2–MC5) in fresh MethoCult M3434 with DMSO or test compounds (n = 2, represented by each circle). D The cellular growth and colony formation activity (MethoCult M3434 assay) of MLL-AF9–evoked murine AML-like cells were monitored after washout of DS-1594a·HCl or Ara-C pretreatment for 7 days. Ara-C, cytarabine