Negative regulation of peroxiredoxin 6 (Prdx 6) transcription by nuclear oncoprotein DEK during leukemia cell differentiation

Figures & data

Figure 1. DEK represses Prdx 6 during leukemia cell differentiation. (A) The expression levels of DEK and Prdx 6 in 293T cells with transiently overexpressed DEK were detected via RT-PCR. The error bars represent 2^−ΔΔC_T ± the SD of three independent experiments. ***P < 0.001. Total proteins were extracts from DEK transfected 293T cells and then were subjected to Western blot analysis using specific antibodies. (B) HL-60 cells were treated with TPA or DMSO. After 48 h, RT-PCR was performed to compare the expression levels of the target genes. Results are shown as means ± SDs; n = 3. **P < 0.01; *P < 0.05. (C) HL-60 cells were treated with ATRA or TPA. After 48 h, differentiated HL-60 cells were lysed and immunoblotted with anti-DEK, anti-β-actin, and anti-Prdx 6 antibodies.

Figure 2. DEK negatively regulates Prdx 6 transcription during leukemia cell differentiation. (A) K562 cells were transfected with Prdx 6-luc and hemin (30 µM) was treated for indicate time points. Cell extracts were assayed for luciferase activity. Luciferase activities were normalized to those of β-galactosidase. Results are shown as means ± SDs; n = 3. **P < 0.01; ***P < 0.001. (B) K562 cells were transfected with Prdx 6-luc and shDEK. Hemin (30 µM) was treated for indicated time points. Cell extracts were assayed for luciferase activity. Luciferase activities were normalized to those of β-galactosidase. Results are shown as means ± SDs; n = 3. **P < 0.01; *P < 0.05.

Figure 3. Occupancy of DEK and the p65 subunit in the Prdx 6 promoter region during leukemia cell differentiation. (A) Schematic diagram of the primer pairs used for ChIP analysis. Black boxes represent the Prdx 6 exons. (B) ChIP analyses of the Prdx 6 promoter in ATRA, TPA-treated HL-60 cells were conducted using anti-DEK, and anti-Prdx 6 and examined via RT-PCR. The results are representative of at least three independent experiments (± SD). ***P < 0.001. *P < 0.05.