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. 2011 Mar 22;286(19):16967–16975. doi: 10.1074/jbc.M111.218206

FIGURE 4.

FIGURE 4.

The transcription factor NF-κB is involved in Bclaf1 transcription in T cells. A and B, luciferase (Luc) plasmids that carry bclaf1 promoter regions were co-transfected with control TK-luciferase plasmids. The luciferase activity in transfected cells was analyzed. -Fold changes when compared with control TK-luciferase activities are shown (A). Bclaf1-luciferase plasmids in A were co-transfected with or without Rel-A expression plasmid. The -fold changes of luciferase activity using different plasmids were normalized to TK-luciferase activities (B). C, the binding sites of NF-κB and E2F1 in bclaf1 promoter were mutated, and their luciferase activities were analyzed in transiently transfected HEK293 cells. D, primary CD4 T cells from Sirt1+/+ and Sirt1−/− mice were stimulated with anti-CD3 or with anti-CD3 plus anti-CD28 for 16 h in the presence or absence of 20 μm NF-κB inhibitor JSH-23. Bclaf1 mRNA levels were analyzed by real-time PCR. E, Sirt1 expression plasmids were co-transfected with Bclaf1-luc and Rel-A expression plasmids. The luciferase activities in transfected cells were analyzed. Error bars represent data (mean ± S.D.) from three independent experiments. Student's t test was used for statistic analysis, NS, no significant difference. **, p < 0.01; ***, p < 0.005.