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. 2002 Sep;22(18):6542–6552. doi: 10.1128/MCB.22.18.6542-6552.2002

FIG. 3.

FIG. 3.

Mapping of transcriptional activation domains in AFX. Schematic diagram (top left) illustrates the conserved domains of AFX. The arrow indicates the fusion site with MLL in human leukemias. Thick horizontal black lines with amino acid numbers represent the AFX fragments fused to the Gal4-DBD for transactivation assays. ENL-C (477 to 559 aa) and ELL-C (496 to 621 aa) correspond to the minimal regions sufficient for immortalization of myeloid progenitors by MLL-ENL and MLL-ELL fusion proteins, respectively (15, 50). VP16-MTD encodes the minimal transcriptional activation domain (aa 413 to 453) of herpes simplex virus VP-16. Expression constructs were cotransfected with pcDNA-LacZ into 293 cells with a luciferase reporter gene under the control of the indicated promoters. TK, herpes simplex virus TK promoter; E1b, adenovirus E1b promoter; Myelomono, myelomonocytic growth factor promoter (described in Materials and Methods). Luciferase values were normalized for β-galactosidase expression from the internal LacZ control construct. Similar expression level of the Gal4 constructs was confirmed by Western blot (data not shown).