Effects of prenyltransferase inhibitors on estrogen response element-dependent luciferase activity in HELN cells, transfected with estrogen receptor (ER) α or its mutants

<p><b>Copyright information:</b></p><p>Taken from "Prenylation inhibitors stimulate both estrogen receptor α transcriptional activity through AF-1 and AF-2 and estrogen receptor β transcriptional activity"</p><p>Breast Cancer Research 2004;7(1):R60-R70.</p><p>Published online 8 Nov 2004</p><p>PMCID:PMC1064103.</p><p>Copyright © 2004 Cestac et al., licensee BioMed Central Ltd.</p> Cells, deprived of estradiol (E2) for 4 days, were co-transfected with the Renilla luciferase plasmid and either HEG0 (full-length ERα), HEG19 (AF-1-deleted ERα), HEG7 (AF-2-deleted ERα) or pSG5 (empty vector). Five hours after transfection, cells were treated or not with FTI-277 (10 μM or dithiothreitol/dimethylsulfoxide vehicle), and 24 hours later they were stimulated with E2 (5 nM) or ethanol and treated or not with FTI-277 or GGTI-298 (10 μM or 5 μM, respectively, or dithiothreitol/dimethylsulfoxide vehicle). Luciferase activity was quantified 16 hours after E2 addition, as described in Materials and methods. Results are expressed in arbitrary units after normalization. Error bars indicate the mean values ± standard deviation from triplicate experiments, and the results are representative of three independent experiments. Results obtained show that prenylation inhibitors statistically increase or do not statistically increase the luciferase activity on their own compared with control cells (white bars) and statistically increase the luciferase activity induced by E2 compared with the activity induced by E2 alone (grey bars) in HELN cells transfected with ERα or its mutants (* < 0.02)

Immunocytochemical analysis of the effects of prenyltransferase inhibitors on estrogen receptor (ER) α distribution in MCF-7 cells

<p><b>Copyright information:</b></p><p>Taken from "Prenylation inhibitors stimulate both estrogen receptor α transcriptional activity through AF-1 and AF-2 and estrogen receptor β transcriptional activity"</p><p>Breast Cancer Research 2004;7(1):R60-R70.</p><p>Published online 8 Nov 2004</p><p>PMCID:PMC1064103.</p><p>Copyright © 2004 Cestac et al., licensee BioMed Central Ltd.</p> Cells, deprived of estradiol (E2) for 7 days, were treated or not with FTI-277 (10 μM or dithiothreitol/dimethylsulfoxide vehicle), and 24 hours later they were stimulated with E2 (5 nM) or ethanol and treated or not with FTI-277 or GGTI-298 (10 μM or 5 μM, respectively, or vehicle). After 24 hours, cells were fixed and stained. One randomly selected field is presented for each treatment. Data were quantified by determining the grey value of both the nucleus and the cytoplasm for each cell counted, as described in Materials and methods. For each experimental condition, six randomly selected fields were analyzed. The total number of cells present in the fields ranged from 250 to 400. Error bars indicate the mean values ± standard error of the mean, and the results are representative of two independent experiments