Co-repressors do not rescue PLZF from inhibition by AML-1/ETO.

293T cells were transiently transfected in 12-well dishes and subjected to luciferase reporter assays. All results are expressed as fold repression compared to the control plasmids. (A) Assays performed with the IL-3Rα-tk-Luc reporter. Lanes 1, 6, 11: pCDNA 400 ng. Lanes 2, 7, 12: PLZF 400 ng. Lanes 3, 8, 13: PLZF 400 ng plus AML-1/ETO 300 ng. Lane 4: PLZF 400 ng and SMRT 300 ng. Lane 5: PLZF 400 ng, SMRT 300 ng, and AML-1/ETO 300 ng. Lane 9: PLZF 400 ng and N-CoR 300 ng. Lane 10: PLZF 400 ng, N-CoR 300 ng, and AML-1/ETO 300 ng. Lane 14: PLZF 400 ng plus ETO 300 ng. Lane 15: PLZF 400 ng, ETO 300 ng, and AML-1/ETO 300 ng. Panels B, C and D: Assays performed with the (GAL4)₅-tk-Luc reporter. (B) Lanes 1, 6: GAL4^1–147 400 ng. Lanes 2, 7: GAL4-PLZF 400 ng. Lanes 3, 8: GAL4-PLZF 400 ng and AML-1/ETO 300 ng. Lane 4: GAL4-PLZF 400 ng and SMRT 300 ng. Lane 5: GAL4-PLZF 400 ng, SMRT 300 ng, and AML-1/ETO 300 ng. Lane 9: GAL4-PLZF 400 ng and ETO 300 ng. Lane 10: GAL4-PLZF 400 ng, ETO 300 ng, and AML-1/ETO 300 ng. Panels C and D correspond to experiments performed with GAL4-BTB/POZ and GAL4-RD2, with doses similar to those in B. The results of all graphs reflect the average of multiple experiments ± SEM.