1. Figure 1. Schematic location of the amino acid substitutions and truncations of the androgen receptor. NTD, N-terminal domain; DBD, DNA binding domain; LBD, ligand binding domain; CAIS, complete androgen insensitivity syndrome; PAIS, partial androgen insensitivity syndrome; MBC, male breast cancer. Numbering is according to Lubahn et al. (47 ). Inset, Immunoblot analysis of the wild-type AR and the different mutants expressed in COS-7 cells and immunostained with antibody SpO61.
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society

2. Figure 2. A, Transactivation of the MVDP-Luc target gene in absence or presence of TPA in CV-1 cells. Transactivation was tested in CV-1 cells by cotransfecting the reporter gene MVDP-Luc with indicated amount of expression vector. Twenty, 50, 100 ng of AR expression vector and 1.5 μg of MVDP-Luc were used. Sixteen hours after transfection, the cells were incubated for 30 h in fresh DMEM medium without serum, in presence of 0.1 or 10 nm R1881. Ten hours before harvesting, 50 nm TPA was added. The total amount by well was kept constant by adding empty pCMV as needed. Presented as a bar diagram are the averages of the relative light units (RLU) with standard deviations from three independent experiments in duplicate. B, Repression of the TRE₃-tk-Luc target genes in CV-1 cells. One hundred nanograms of empty vector or AR expression vector and 1.5μ g of TRE₃-tk-Luc were used. Cells were incubated with R nm alone for 30 h, or with TPA 50 nm for 10 h. The results, expressed as induction factors, are the averages with standard deviations from at least three independent experiments in duplicate. In the absence of TPA, the value of induction factor was 1. The baseline was not significantly modified by incubation with androgen alone (less than 10%).
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society

3. Figure 6. Transactivation of MVDP-Luc (A) and transrepression of TRE₃-tk-Luc (B) reporter genes, respectively, in CV-1 cells. One hundred nanograms of AR expression vectors and 1.5 μg of each reporter gene were used. Cells were incubated with R nm for 30 h or TPA 50 nm for 10 h, alone or combined. Presented as a bar diagram are the averages of the induction factors with standard deviations from at least five independent experiments in duplicate. DBD, LBD, and NTD indicate the mutants from the DNA binding, the ligand binding, and the N-terminal domains, respectively; CAIS, complete androgen insensitivity syndrome; PAIS, partial androgen insensitivity syndrome; MBC, male breast cancer.
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society

4. Figure 5. Effects of 30 nm staurosporine, 1 nm calyculin-A, and 1 nm okadaic acid on R1881-induced luciferase activity in CV-1 cells. Transfected cells with 1.5 μg of MVDP-Luc and 100 ng of AR were treated for 20 h with or without 10 nm R1881 and then incubated for another 10 h with the various compounds indicated in the figure. Results are expressed as the percentage of luciferase activity in CV-1 cells from at least three independent experiments in duplicate. The 100% and 0% values were obtained with 10 nm R1881 and ethanol, respectively. Effects of staurosporine, calyculin-A and okadaic acid were compared with that of TPA alone. Student’s t test was used for statistical analysis. ***, P = 0.001;* , P = 0.01.
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society

5. Figure 4. A, Androgen-regulated luciferase activity in CV-1 cells transfected by c-Fos or c-Jun. Cells were transfected by 100 ng of AR, 1.5 μg of reporter gene MVDP-Luc, and either c-Fos or of c-Jun expression vectors (amount indicated as ng in the figure). The luciferase signal measured in the cells transfected by the empty vector pCI was arbitrarily set at 100%. B, Effects of different cotransfections on basal activity of constitutive gene reporter pCMV-Luc. CV-1 cells were cotransfected with 1.5 μg of pCMV-Luc and expression vectors as follows: empty expression vectors (pCMV: 100 ng and pCI : 400 ng) and expression vectors (AR: 100 ng, c-Fos and c-Jun: 400 ng). Cells were treated with R1881 for 30 h after removal of the calcium precipitate. Columns represent the average of three experiments in duplicate.
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society

6. Figure 3. A, MVDP- and TRE₃-tk- controlled luciferase activity in presence of 50 nm of TPA for different times. To measure inhibition of transactivation in presence of TPA, transfected cells with 100 ng of AR expression vector and 1.5μ g of MVDP-Luc were incubated with R nm for 30 h combined with TPA for 6, 8, 10, 12, and 24 h. For assays of AP-1 transactivation, cells were transfected with TRE₃-tk-Luc (1.5 μg) and 100 ng of mock pCMV5 vector and incubated with R nm and TPA as described above. B, CV-1 cells were transfected with TRE₃-tk-Luc or MVDP-Luc as described above and incubated with R nm for 30 h and combined for 10 h with various amounts of TPA (0.1, 0.5, 1, 5, 10, and 50 nm). In both figures, the transactivation of the two reporter genes is expressed as fold-induction of luciferase activity. Each data point was tested in duplicate and induction of luciferase activity was calculated from three different experiments.
Transcriptional Interferences between Normal or Mutant Androgen Receptors and the Activator Protein 1—Dissection of the Androgen Receptor Functional Domains**Part of this work was presented at the 10th International Congress of Endocrinology, San Francisco, California, June 12–15, 1996 and was awarded the Henning Andersen Prize (best abstract) at the 35th Annual Scientific Meeting of the European Society for Pædiatric Endocrinology, Montpellier, France, September 15–18, This work was supported by the Institut National de la Santé et de la Recherche Médicale; the Université Montpellier I; the Association pour la Recherche sur le Cancer, Grant 6711; and the Fédération Nationale des Centres de Lutte contre le Cancer, Grant
Endocrinology. 1999;140(1): doi: /endo
Endocrinology | Copyright © 1999 by The Endocrine Society