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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Leukemia. 2012 Nov 9;27(4):813–822. doi: 10.1038/leu.2012.327

Fig.3. EPZ004777 selectively inhibits proliferation of MLL-AF10 and CALM-AF10 transformed murine bone marrow cells.

Fig.3

(A) Growth of MLL-AF10, CALM-AF10, MLL-AF9, and Hoxa9/Meis1a transformed bone marrow cells during several days’ incubation with 10 µM EPZ004777. Viable cells were counted and replated at equal cell numbers in fresh media with fresh compound every 3–4 days. Results were plotted as percentage of split-adjusted viable cells in the presence of 10 µM EPZ004777 compared to DMSO vehicle control. Results are representative of three independent experiments.

(B) Dosage effect of EPZ004777 treatment on MLL-AF10, CALM-AF10, MLL-AF9, and HoxA9/Meis1a transformed bone marrow cells. Cells were counted and replated at equal cell numbers in fresh media with fresh compound every 3–4 days. Results were plotted as percentage of split-adjusted viable cells on day 17 in media with 0.1 µM, 1 µM, 10 µM of EPZ004777 compared to DMSO control (set as 100%). Results are representative of three independent experiments.

(C) Time course of Hoxa9 and Meis1 mRNA expression in MLL-AF10 and CALM-AF10 transformed cells over 10 days of incubation with 10 µM EPZ004777 as measured by quantitative real-time PCR. Expression levels were normalized to Gapdh and expressed relative to those at day 0 (set to 100%). Error bars indicate the SEM (n=3 independent experiments).

(D) Quantitative real-time PCR analysis of Hoxa9, Meis1, and β-Actin mRNA levels in MLL-AF10 and CALM-AF10 transformed cells following 7 days of incubation with EPZ004777. Relative mRNA expression levels are plotted as a percentage of those in vehicle-treated control cells. Error bars represent SEM (n=3 independent experiments).

(E) Inhibition of cellular H3K79me2 levels in MLL-AF10, CALM-AF10, MLL-AF9 or Hoxa9-Meis1a transformed bone marrow cells following 7 days of treatment with the indicated concentrations of EPZ004777 as measured by immunoblot analysis of extracted histones with an anti-H3K79me2 antibody.