FIGURE 5.

Ets-2 is a general repressor of IL-2. A, Ets-2 and IL-2 mRNA levels in different leukemic cell lines measured by RT-PCR. All cells were cultured in CM. Reverse transcribed β2-m from each cell line was used as an internal control. B, Ets-2 and IL-2 mRNA levels were measured by real time PCR in Raji cells transfected with varying amounts of pcDNA3-ets-2 plasmid (0, 10, 20, and 30 μg). The amount of transfected DNA/sample was retained to 60 μg by the addition of appropriate amounts of vector pcDNA3 DNA. Cells were kept in CM after transfection for 6 h. β2-m was used as an internal control. Error bars represent S.E. (*, p < 0.05; **, p < 0.01, two-way ANOVA). C, Western blotting analysis to confirm overexpression of Ets-2 protein in transfected Raji cells. Raji cells were transfected with 20 μg of pcDNA3-ets-2 DNA or pcDNA3 and lysed 6 h post-transfection, and total protein extracts were prepared. Tubulin was used as an internal control for equal loading. The results shown in A, B, and C are representative of three independent experiments.