Fig. 2
SHARP1 is a downstream target of MLL-AF6 and DOT1L. a SHARP1 mRNA expression in human AML cell lines assessed by qPCR. The cell lines analyzed are: ML-2 (MLL-AF6), CTS (MLL-AF6), SHI-1 (MLL-AF6), MOLM-14 (MLL-AF9), MV4-11 (MLL-AF4), and Kasumi-1 (AML1-ETO). b ChIP-seq profiles of ML-2 cells using MLLN, MEN1, LEDGF, H3K79me2, and H3K79me3 antibodies at the loci of the HOXA gene cluster (left panel) and SHARP1 gene (right panel). c ChIP-seq profiles of SHI-1 cells using MLLN antibody and input at the loci of the HOXA gene cluster (left) and SHARP1 gene (right). d qPCR for MLL-AF6 (left panel) and SHARP1 (right panel) mRNA expression in ML-2 cells upon MLL knockdown. Shown is the relative expression value to ML-2 transduced with shGFP. e Western blot of H3K79me2 in MOLM-14 (MLL-AF9) and ML-2 (MLL-AF6) cells treated with the DOT1L inhibitor, EPZ5679 (1 μM) or DMSO vehicle for 96 h. f qPCR for SHARP1 mRNA in MOLM-14 or ML-2 cells treated with EPZ5679 or DMSO vehicle for 6 days. Relative expression is the value compared to ML-2 cells treated with DMSO vehicle. g Western blot of SHARP1 and β-actin in ML-2 cells treated with EPZ5679 or DMSO vehicle for 10 days. All Western blots are representative of three independent experiments. All quantitation data include three independent experiments and are presented as mean ± s.e.m
