A complex containing LMO2, Lyl1, LDB1, SP1, and HEB or E2A activates the DLEU2 promoter. (A) DLEU2 promoter sequence from position −733 to −378 that contains an SP1 and an E2A binding sites. The E-box and GC-box are indicated in bold and underlined. (B) LMO2 protein indirectly associates with the DLEU2 promoter in vivo. Chromatin extracts from the OCILy1 cell line induced to express LMO2 protein were subjected to immunoprecipitation with FLAG (LMO2) and GAPDH antibodies and control IgG. Enriched DNA fragments were quantified by real-time PCR using. The GAPDH gene was used as a negative control. The fold enrichment was expressed as the percentage of input. Results are representative of 3 independent experiments. (C) DLEU2 promoter region (A) was cloned in a pGL3 plasmid. Raji cells were transfected with the DLEU2 Luciferase promoter (10 μg), internal control plasmid pRLTK (10 ng), indicated expression vectors (E12, E47, HEB, SP1, LMO2, Lyl1, LDB1, 2 μg each) and control pCDNA3.1 plasmid that was used to keep constant the total amount of transfected DNA. Luciferase activity was determined 48 hours after transfection. The values are relative luciferase activities, with the average value obtained for the same amount of reporter and pcDNA3.1 control plasmid set at 1. Values are means + SE of 3 independent experiments, each performed in 3 replicate samples.