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. 2009 Jun 4;284(31):20593–20601. doi: 10.1074/jbc.M109.021477

FIGURE 2.

FIGURE 2.

DLX5 regulates MYC transcription in vitro. A, annotation of the MYC promoter. SNM and XNM, sequences inserted in SNM-Luc and XNM-Luc reporter vectors, respectively. The MYC promoter contains four putative DLX5 binding sites with the core motif of TAAT. P1 and P2 sites are major transcription start sites. B and C, luciferase activity in cells transfected with XNM-Luc or SNM-Luc, respectively, normalized by Renilla luciferase. pcDNA3.1 (Vec) in samples 1 and 3 or pcDNA3.1-DLX5 (DLX5) in samples 2 and 4 was co-transfected with luciferase reporter constructs and Renilla luciferase plasmid. Statistical analysis was performed by a two-tailed t test. D, MYC promoter reporter constructs bearing four different DLX5 binding sites were subjected to site-directed mutagenesis to substitute putative DLX5 binding sequences with the designated sequences (underlined). E, promoter activity measured using XNM-Luc or XNM-M4-Luc (Xmut-Luc), which contains a mutation in the D4 binding site of DLX5 in HeLa cells. Luciferase activity was normalized by Renilla luciferase. F, promoter activity of four different SNM-Luc mutant vectors (M1-Luc, M2-Luc, M3-Luc, and M4-Luc) containing different mutations in putative DLX5 binding sites in SNM-Luc in HeLa cells. Luciferase activity was normalized by Renilla luciferase.