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| Status |
Public on Jun 20, 2005 |
| Title |
Novel N-terminal Glucocorticoid Receptor Isoforms with Unique Transcriptional Target Genes |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Gene expression analysis was conducted using Agilent Human1Av2 arrays (Agilent Technologies, Palo Alto, CA). Two separate biological replicates of cytoplasmic RNA samples were harvested and purified from each of the U-2 OS cell lines stably expressing hGRalpha, -A, -B, -C3, or D3 treated with 100 nM DEX or vehicle for 6 hours using RNeasy Midi kits (Invitrogen). These RNA samples were amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 0.5 ng of total RNA, two cRNAs, one labeled with Cy3 and one labeled with Cy5 were produced for each sample according to the manufacturer’s protocol. Consequently, four separate chips were used for each of the isoforms at each treatment condition, yielding a total of 40 chips: 5 isoforms X 2 treatments (vehicle vs. hormone) X 2 biological replicates X 2 labeling dyes. For each two-color comparison, 750 ng of each Cy3 and Cy5 labeled cRNAs were mixed and fragmented using the Agilent In Situ Hybridization Kit protocol. Hybridizations were performed for 16 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 were obtained using the Agilent Feature Extraction software (v7.1), using defaults for all parameters.
Keywords: other
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| Contributor(s) |
Lu NZ, Cidlowski JA |
| Citation(s) |
15866175 |
| Submission date |
Mar 22, 2005 |
| 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 (20)
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| GSM45610 |
hGRalpha-A vehicle vs. hGRalpha-A DEX, Array 1, Biorep 1 |
| GSM45611 |
hGRalpha-A vehicle vs. hGRalpha-A DEX, Array 2, Biorep 1 |
| GSM45614 |
hGRalpha-B vehicle vs. hGRalpha-B DEX, Array 1, Biorep 1 |
| GSM45615 |
hGRalpha-B vehicle vs. hGRalpha-B DEX, Array 2, Biorep 1 |
| GSM45651 |
hGRalpha vehicle vs. hGRalpha DEX, Array 1, Biorep 1 |
| GSM45652 |
hGRalpha vehicle vs. hGRalpha DEX, Array 2, Biorep 1 |
| GSM45653 |
hGRalpha-D3 vehicle vs. hGRalpha-D3 DEX, Array 1, Biorep 1 |
| GSM45654 |
hGRalpha-D3 vehicle vs. hGRalpha-D3 DEX, Array 2, Biorep 1 |
| GSM45655 |
hGRalpha-C3 vehicle vs. hGRalpha-C3 DEX, Array 1, Biorep 1 |
| GSM45656 |
hGRalpha-C3 vehicle vs. hGRalpha-C3 DEX, Array 2, Biorep 1 |
| GSM45665 |
hGRalpha vehicle vs. hGRalpha DEX, Array 1, Biorep 2 |
| GSM45666 |
hGRalpha vehicle vs. hGRalpha DEX, Array 2, Biorep 2 |
| GSM45667 |
hGRalpha-C3 vehicle vs. hGRalpha-C3 DEX, Array 1, Biorep 2 |
| GSM45668 |
hGRalpha-C3 vehicle vs. hGRalpha-C3 DEX, Array 2, Biorep 2 |
| GSM45669 |
hGRalpha-D3 vehicle vs. hGRalpha-D3 DEX, Array 1, Biorep 2 |
| GSM45670 |
hGRalpha-D3 vehicle vs. hGRalpha-D3 DEX, Array 2, Biorep 2 |
| GSM45671 |
hGRalpha-A vehicle vs. hGRalpha-A DEX, Array 1, Biorep 2 |
| GSM45672 |
hGRalpha-A vehicle vs. hGRalpha-A DEX, Array 2, Biorep 2 |
| GSM45673 |
hGRalpha-B vehicle vs. hGRalpha-B DEX, Array 1, Biorep 2 |
| GSM45674 |
hGRalpha-B vehicle vs. hGRalpha-B DEX, Array 2, Biorep 2 |
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| Relations |
| BioProject |
PRJNA91587 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE2428_RAW.tar |
613.3 Mb |
(http)(custom) |
TAR (of TIFF, TXT) |
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