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. 2016 Feb 29;126(4):1438–1450. doi: 10.1172/JCI80825

Figure 2. Loss of Dot1l leads to decreased growth, increased differentiation, and apoptosis of MN1-driven CMP-derived murine leukemia cells.

Figure 2

(A) Serial replating of MN1-transformed CMPs (MN1CMP-T) after Cre-induced loss of Dot1l. Left plot: number of colonies per 500 plated cells; right plot: total cell number. n = 5 independent experiments; two-sided t test Dot1l–/– vs. Dot1lfl/fl. Error bars represent ±SEM. (B) Serial replating of MN1-driven murine leukemias (MN1CMP-L) after Cre-induced loss of Dot1l. Left plot: number of colonies per 500 plated cells; right plot: total cell number. Dot1lfl/fl: bulk population from 3 independent experiments; Dot1l–/–: 2 bulk populations and 4 individually picked clones (due to outgrowth of nondeleted cells) from 3 independent experiments. Two-sided t test Dot1l–/– vs. Dot1lfl/fl. Error bars represent ±SEM. (C) Methylcellulose colony and leukemia cell morphology (Wright-Giemsa stain) of MN1-driven murine leukemias (MN1CMP-L) 14 days after transduction with Cre. Representative of 3 independent expermients. (D) CD11b expression in MN1-driven murine leukemias (MN1CMP-L) 3 weeks after deletion of Dot1l. Dot1lfl/fl: bulk population from 3 independent experiments; Dot1l–/–: 2 bulk populations and 4 individually picked clones (due to outgrowth of nondeleted cells) from 3 independent experiments. Two-sided t test Dot1l–/– vs. Dot1lfl/fl. Error bars represent ±SEM. (E) Apoptosis (Annexin staining) in MN1-driven murine leukemias (MN1CMP-L) 3 weeks after deletion of Dot1l. Dot1lfl/fl: bulk population from 3 independent experiments; Dot1l–/–: 2 bulk population and 4 individually picked clones (due to outgrowth of nondeleted cells) from 3 independent experiments. Two-sided t test Dot1l–/– vs. Dot1lfl/fl. Error bars represent ±SEM. (F) Cell cycle distribution (EdU incorporation/DAPI staining) in MN1-driven murine leukemias (MN1CMP-L) 3 weeks after deletion of Dot1l. Dot1lfl/fl: bulk population from 3 independent experiments; Dot1l–/–: 2 bulk population and 4 individually picked clones (due to outgrowth of nondeleted cells) from 3 independent experiments. Two-sided t test Dot1l–/– vs. Dot1lfl/fl. Error bars represent ±SEM.