American Society of Hematology

Figure 2

$Figure 2. The ID1 promoter is transactivated by PML-RARα but not by RARα/RXR. (A) Hep3B cells were transfected with the 963-bp ID1 promoter-luciferase reporter construct (ID1-luc) together with a control vector expressing Renilla luciferase. In addition, vectors coding for the various proteins indicated in the figure were transfected. Transactivation is expressed as arbitrary units and is corrected for transfection efficiency measured by Renilla luciferase. Background luminescence of the cells transfected with only the reporter construct without nuclear receptors and in the absence of ATRA was set at 1. Cells were cultured for 16 hours without (▭) and with () ATRA. Mean values and standard deviations from 3 independent experiments (± SD) are shown. (B) To show transactivation by unrearranged retinoic acid receptors, cells were transfected with the RARE3-tk-luc vector, containing 3 bona fide RAREs. Mean values of 3 independent experiments (± SD) are shown. (C) ID1 expression is increased by PML-RARα in the presence of ATRA. The zinc-inducible PML-RARα cell line U937-PR9 was grown in the absence (left panel) or presence (right panel) of zinc for 16 hours. Cells were treated with ATRA and harvested at the indicated time points. Proteins were size-fractionated by SDS–polyacrylamide gel electrophoresis (SDS-PAGE). Immunostaining was done with anti-ID1 antibody.$

The ID1 promoter is transactivated by PML-RARα but not by RARα/RXR. (A) Hep3B cells were transfected with the 963-bp ID1 promoter-luciferase reporter construct (ID1-luc) together with a control vector expressing Renilla luciferase. In addition, vectors coding for the various proteins indicated in the figure were transfected. Transactivation is expressed as arbitrary units and is corrected for transfection efficiency measured by Renilla luciferase. Background luminescence of the cells transfected with only the reporter construct without nuclear receptors and in the absence of ATRA was set at 1. Cells were cultured for 16 hours without (▭) and with () ATRA. Mean values and standard deviations from 3 independent experiments (± SD) are shown. (B) To show transactivation by unrearranged retinoic acid receptors, cells were transfected with the RARE3-tk-luc vector, containing 3 bona fide RAREs. Mean values of 3 independent experiments (± SD) are shown. (C) ID1 expression is increased by PML-RARα in the presence of ATRA. The zinc-inducible PML-RARα cell line U937-PR9 was grown in the absence (left panel) or presence (right panel) of zinc for 16 hours. Cells were treated with ATRA and harvested at the indicated time points. Proteins were size-fractionated by SDS–polyacrylamide gel electrophoresis (SDS-PAGE). Immunostaining was done with anti-ID1 antibody.

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