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. 2009 Dec 14;285(8):5594–5605. doi: 10.1074/jbc.M109.068346

FIGURE 3.

FIGURE 3.

GILZ inhibits FOXO4-WT transcriptional activity in HL-60 cells. HL-60 cells were transiently transfected with 10 μg of pMT2-FOXO4-WT and/or 10 μg of pcDNA3-Myc-GILZ or pcDNA3-Myc and with either 10 μg of the reporter plasmid pBim-Luc (A) or 10 μg of p27-Luc (B) or pFasL-Luc (C). The transcriptional activity was analyzed after 24 h of culture. Results are expressed as the percentage of reporter activity, with 100% representing the activity of the construct in the presence of FOXO4-WT. Data represent the mean ± S.E.M. of 3 independent experiments performed in triplicate. *, p < 0.05. D, HL-60 cells were transiently transfected with the reporter plasmid p3xIRS-MLP-Luc and either 1 μg of pMT2-FOXO4-WT and/or 5 μg of pcDNA3-Myc-GILZ or pcDNA3-Myc. Results are expressed as the percentage of p3xIRS-MLP-Luc activity, with 100% representing the activity of the construct in the presence of FOXO4-WT. Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. *, p < 0.05. E, HL-60 cells were transiently transfected with 10 μg of pcDNA3-FOXO4-WT and/or 10 μg of pcDNA3-Myc-GILZ or pcDNA3-Myc, and cells were harvested after 24 h of culture. Western blot was performed using anti-FOXO4 and anti-GILZ antibodies. β-Tubulin was used as an internal control for protein levels. IB, immunoblot. Error bars, S.E.