FIGURE 3.

Effect of E2A-HLF on survivin promoter activity in transiently transfected t(17;19)⁻ ALL cells. A, Nalm-6/E2A-HLF cells cotransfected with pRL-TK vector and the pGL3-survivin promoter constructs indicated at the left were cultured in the absence (open bars) or presence (black bars) of zinc for 24 h. Firefly luciferase (Luc) activity was normalized to Renilla luciferase as a transfection efficiency control. The level of activity of the promoterless Renilla plasmid luciferase was defined as 1. The results depicted are the averages of three independent experiments; error bars indicate S.D. # indicates mutation of CHR, and ** indicates mutation of CDE. B, nucleotide sequences of the human survivin promoter and three mutants. Underlines indicate CHR or CDE region. Shaded characters indicate mutation (mut). C, EMSA. Nuclear lysates extracted from Nalm-6/E2A-HLF cells cultured without (lanes 1 and 3–5) or with zinc (lane 2) were incubated with a ³²P-end-labeled oligonucleotide probe containing the CHR sequence (lanes 1–4) or mutated CHR sequence (lane 5). An excess of unlabeled CHR sequence competitor (lane 3) or mutant competitor (lane 4) was added to the reaction mixture. Mt, mutant.