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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Nature. 2010 Jul 18;466(7307):765–768. doi: 10.1038/nature09171

Figure 2. The RNA binding protein Musashi is highly expressed in immature normal and leukemic cells and is regulated by HoxA9.

Figure 2

a, Musashi (Msi) expression in whole bone marrow (WBM), KLS cells, chronic and blast crisis CML, olfactory bulb (OB), –reverse transcriptase (-RT in OB) and water. Tbp, TATA binding protein. b–e, Realtime RT-PCR analysis of Msi2 expression in b, KLS cells (n=3), Lin+ cells (n=2) **p<0.001, c, blast crisis phase (n=9), chronic phase (n=6) **p<0.001, d, lin− chronic and blast crisis phase cells relative to normal KLS and lin+ cells (lin+, n=2 and others, n=3), e, lin− (n=5) or lin+ (n=5) blast crisis CML cells *p=0.039. Error bars represent s.e.m. f, Control vector- or Msi2-expressing CML cells were stained with anti-Numb antibody (red) and DAPI (green pseudocolor) and g, fluorescence intensity quantified **p<0.001. h, Msi2 expression in KLS cells transduced with either control vector or NUP98-HOXA9 retrovirus along with BCR-ABL *p=0.017. i–l, HoxA9 binds to the Msi2 promoter. Murine Msi2 gene structure: exons (numbered boxes), transcription start site (TSS; +1) and the direction of transcription (flag), putative HOX binding element 5.7kb upstream of TSS (oval) and +110kb site with no HoxA9 binding sequence (open rectangle). i, ChIP was performed either with IgG control or anti-HoxA9 antibody for j, Flt3, a known HoxA9 target gene, as a positive control and k, Msi2 −5.7kb region or l, Msi2 +110kb region. m, KLS cells from Msi2 genetrap reporter mice were transduced with BCR-ABL with either control vector or NUP98-HOXA9 and β-galactosidase reporter activity quantified (n=2 each, *p=0.011).