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| Status |
Public on Mar 02, 2007 |
| Title |
hGRβ: Elucidation of its Role in Transcriptional Regulation and Identification of a Ligand |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
The human glucocorticoid receptor (GR) is expressed as two alternately spliced Cterminal isoforms: α and β. In contrast to the canonical hGRα, hGRβ is constitutively nuclear, is not thought to bind ligand, and is believed to affect gene transcription only by acting as a dominant negative to hGRα. Microarray analysis indicated that hGRβ, expressed in the absence of hGRα, can regulate gene expression, and furthermore, occupation of hGRβ with the antagonist RU-486 diminishes that capacity.
Keywords: cell type comparison
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| Overall design |
RNA preparation: Total RNA was extracted from 5x10 6 U-OFF or U-2
OSβ cells treated with either ethanol vehicle or 1μM RU-486 for 6 hours using the RNAqueous Total RNA Isolation Kit (Ambion Inc. Austin, TX) according to manufacturer’s instructions. RNA was treated with DNase using the DNA-Free DNase Treatment and Removal Reagents (Ambion Inc.) according to manufacturer’s instructions prior to use with the microarray. Four pairs of RNA (vehicle vs. RU-486 treated) were harvested for each cell type to yield four biological replicates for gene expression analysis.
Linear Amplification Label Protocol and Feature Extraction: Gene expression analysis was conducted using Agilent Human1Av2 arrays (Agilent Technologies, Palo Alto, CA). Total RNA was amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 500ng of total RNA, Cy3 or Cy5 labeled cRNA was produced according to manufacturer’s protocol. For each two color comparison, 750ng of each Cy3 and Cy5 labeled cRNAs were mixed and fragmented using the Agilent In Situ Hybridization Kit protocol. Hybridizations were performed for 17 hours in a rotating hybridization oven using the Agilent 60-mer oligo microarray processing protocol. Slides were washed as indicated in this protocol
and then scanned with an Agilent Scanner. Data was obtained using the Agilent Feature Extraction software (v7.5), using defaults for all parameters.
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| Contributor(s) |
Lewis-Tuffin LJ, Jewell CM, Bienstock RJ, Collins JB, Cidlowski JA |
| Citation(s) |
17242213 |
| Submission date |
Jul 14, 2006 |
| Last update date |
Dec 06, 2012 |
| Contact name |
NIEHS Microarray Core |
| E-mail(s) |
microarray@niehs.nih.gov, liuliw@niehs.nih.gov
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| Organization name |
NIEHS
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| Department |
DIR
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| Lab |
Microarray Core
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| Street address |
111 T.W. Alexander Drive
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| City |
RTP |
| State/province |
NC |
| ZIP/Postal code |
27709 |
| Country |
USA |
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| Platforms (1) |
| GPL887 |
Agilent-012097 Human 1A Microarray (V2) G4110B (Feature Number version) |
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| Samples (24)
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| GSM119878 |
hGRB con 1/27/05 vs. hGRB RU 1/27/05 (ExpID 2800) |
| GSM119879 |
hGRB RU 1/27/05 vs. hGRB con 1/27/05 (ExpID 2800) |
| GSM119880 |
hGRB con 2/7/05 vs. hGRB RU 2/7/05 (ExpID 2801) |
| GSM119881 |
hGRB RU 2/7/05 vs. hGRB con 2/7/05 (ExpID 2801) |
| GSM119882 |
hGRB con 2/16/05 vs. hGRB RU 2/16/05 (ExpID 2802) |
| GSM119883 |
hGRB RU 2/16/05 vs. hGRB con 2/16/05 (ExpID 2802) |
| GSM119884 |
hGRB con 2/23/05 vs. hGRB RU 2/23/05 (ExpID 2803) |
| GSM119885 |
hGRB RU 2/23/05 vs. hGRB con 2/23/05 (ExpID 2803) |
| GSM119886 |
U-off con 1/26/05 vs. U-off RU 1/26/05 (ExpID 2793) |
| GSM119887 |
U-off RU 1/26/05 vs. U-off con 1/26/05 (ExpID 2793) |
| GSM119888 |
U-off con 2/3/05 vs. U-off RU 2/3/05 (ExpID 2795) |
| GSM119889 |
U-off RU 2/3/05 vs. U-off con 2/3/05 (ExpID 2795) |
| GSM119890 |
U-off con 2/8/05 vs. U-off RU 2/8/05 (ExpID 2797) |
| GSM119891 |
U-off RU 2/8/05 vs. U-off con 2/8/05 (ExpID 2797) |
| GSM119892 |
U-off con 2/10/05 vs. U-off RU 2/10/05 (ExpID 2799) |
| GSM119893 |
U-off RU 2/10/05 vs. U-off con 2/10/05 (ExpID 2799) |
| GSM119895 |
U-off con 1/26/05 vs. hGRB con 1/27/05 (ExpID 2831) |
| GSM119896 |
hGRB con 1/27/05 vs. U-off con 1/26/05 (ExpID 2831) |
| GSM119897 |
U-off con 2/3/05 vs. hGRB con 2/7/05 (ExpID 2832) |
| GSM119898 |
hGRB con 2/7/05 vs. U-off con 2/3/05 (ExpID 2832) |
| GSM119900 |
U-off con 2/8/05 vs. hGRB con 2/16/05 (ExpID 2833) |
| GSM119901 |
hGRB con 2/16/05 vs. U-off con 2/8/05 (ExpID 2833) |
| GSM119902 |
U-off con 2/10/05 vs. hGRB con 2/23/05 (ExpID 2834) |
| GSM119903 |
hGRB con 2/23/05 vs. U-off con 2/10/05 (ExpID 2834) |
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| Relations |
| BioProject |
PRJNA96319 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE5310_RAW.tar |
739.9 Mb |
(http)(custom) |
TAR (of TIFF, TXT) |
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