Targeting SIRT1 inhibits proliferation and self-renewal in murine leukemia models. (A) Illustration of experimental procedures. (B) Western blot of BM cells from leukemic FLT3^wt/wt-MLL/AF9 (B6-M/A9), FLT3^ITD/ITD-MLL/AF9 (F3-M/A9), AML1-ETO_Mx1-Cre-MSCV-FLT3-ITD (A/E-F3-ITD), and preleukemic AML1-ETO_Mx1-Cre-MSCV-FLT3-WT (A/E-F3-WT) or AML1-ETO_Mx1-Cre-MSCV (A/E-EV) mice probed with Sirt1, phospho-Flt3, Flt3, and Actin-specific antibodies. (C) Quantitative analysis of SIRT1 mRNA levels in leukemic BM cells of the indicated murine leukemia models. SIRT1 expression was normalized to GAPDH expression. Data shown are mean values (n = 3; M/A9 [left], A/E [right]). (D) Serial replating assays of leukemic BM cells derived from the indicated leukemia models, and 1×10⁵ cells were plated in M3434 methylcellulose and treated with vehicle, TV-6, PKC412, or in combination as indicated. After 8 days, cells were pooled and replated at 1×10⁵ cells in each round. Results shown are mean values ± SD of duplicate measurements of 3 independent experiments.