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. 2001 Aug;21(16):5577–5590. doi: 10.1128/MCB.21.16.5577-5590.2001

FIG. 7.

FIG. 7

Transactivation analysis of the Bcl-2 promoter with AML1-ETO. (A) U937T and U937T-A/E cells were transfected with either promoterless firefly luciferase reporter pXP1 or the 3.7-kb Bcl-2 gene upstream sequence–firefly luciferase reporter pBcl2-luc after they had been cultured in the absence or presence of tetracycline for 12 h. The pRL-Null Renilla luciferase construct was cotransfected to normalize transfection efficiency. (B) U937T cells were transfected with pXP1 or pBcl2-luc and various amounts of pCMV5 or pCMV5-AML1-ETO, along with pRL-Null to normalize transfection efficiency. The activity of the promoter was calculated as the ratio of the firefly luciferase activity and the Renilla luciferase activity. The transactivation was calculated as the ratio between pXP1 and pBcl2-luc with 0, 2, and 4 μg of pCMV5-AML1-ETO, assuming a value of 1 for each in the presence of pXP1. (C) U937T cells were transfected with MDR-1-luc and various amounts of pCMV5 or pCMV5-AML1-ETO, along with pRL- Null to normalize transfection efficiency. The activity of the promoter was calculated as the ratio of the firefly luciferase activity and the Renilla luciferase activity. The transactivation was calculated as the ratio between 0 μg and 2 or 4 μg of pCMV5-AML1-ETO. The results are the means of three independent experiments (standard errors of the means are shown when measurable).