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Sample GSM1071268 Query DataSets for GSM1071268
Status Public on Jan 27, 2013
Title AtT20 cell_48hr_HX630 10μM_rep3
Sample type RNA
Source name AtT20 cell, 48hr, HX630 μM, replicate 3
Organism Mus musculus
Characteristics cell line: murine pituitary corticotroph tumor cells (AtT20 cell)
agent: HX630
Treatment protocol AtT20 cells grown to 50 % confluence in regular medium in 12-multiwell plates were incubated either without or with an RXR agonist HX630 at 10 μM in DMEM supplemented with 1 % stripped FBS media for 48 hr.
Growth protocol AtT20 cells were grown with Dulbecco's modified Eagle medium (DMEM) supplemented with 10 % fetal bovine serum (FBS), 100 U/mL penicillin and 100 µg/mL streptomycin. Cells were cultured in a humidified incubator at 37 C with 5 % CO2.
Extracted molecule total RNA
Extraction protocol Total RNAs were isolated using NucleoSpin RNA II (TaKaRa Bio, Ohtsu, Japan) according to the manufacturer’s instructions. Samples were treated with DNase before the experiments in order to remove any contaminant DNA. The RNA was quantified by a Nanodrop 2000 (Thermo Scientific). The quality of the RNA for microarray was evaluated by an Agilent BioAnalyzer 2100. RNA samples with RIN (RNA integrity number) >8.0 and A260/A280 of approximately 2.0 were used for the gene expression analysis.
Label Cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 200 ng RNA using a Low Input Quick-AMP labeling kit (Agilent Technologies) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN). Dye incorporation and cRNA yield were checked with the NanoDrop Spectrophotometer.
Hybridization protocol 1.5 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 25x Agilent fragmentation buffer and 10x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent GE hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Mouse Genome Oligo Microarrays (G4846A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately.
Scan protocol Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5 um, Dye channel is set to Green and Green PMT is set to 100%).
Description Gene expression in AtT20 cells treated with HX630 at 10 μM for 48 hr
Data processing The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters (protocol GE1_107_Sep09 and Grid: 026655_D_F_20100123) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
Advanced Flag Improt Setting:
Feature is not positive and significant :Not detected
Feature is not uniform: compromised
Feature is not above background: Not detected
Feature is Saturated : compromised
Feature is a population outliler?compromised
Normalization Options:
Threshold raw signal to 1.0
Normalization algorithm : Percentile Shift
Percentile target: 75(%)
Baseline Options:
Do not perform baseline transformation.
Submission date Jan 27, 2013
Last update date Jan 30, 2013
Contact name Akiko Saito-Hakoda
Organization name Tohoku University Graduate School of Medicine
Department Molecular Endocrinology
Street address 2-1, Seiryo-machi, Aoba-ku
City Sendai
ZIP/Postal code 980-8575
Country Japan
Platform ID GPL11202
Series (1)
GSE43783 Microarray analysis of an RXR agonist HX630 regulated gene expression in murine pituitary corticotroph tumor cells (AtT20 cell)

Data table header descriptions
VALUE Normalized signal intensity

Data table
GE_BrightCorner 3.0726023
DarkCorner -8.871307
A_55_P1989846 -2.2572756
A_55_P1991598 -6.930971
A_55_P2022211 -8.752296
A_55_P1980764 -2.5776682
A_55_P1964375 1.811635
A_51_P128876 -0.6404228
A_55_P2121042 -8.756777
A_52_P219230 -8.758789
A_51_P207591 -6.933857
A_55_P2131920 -8.76397
A_55_P2404223 -2.0611296
A_55_P2101944 3.786769
A_52_P358860 -0.4193678
A_51_P119031 -0.4006157
A_51_P309854 -3.4861488
A_51_P343900 3.909855
A_51_P234359 2.632185
A_51_P487813 -7.8111753

Total number of rows: 39485

Table truncated, full table size 924 Kbytes.

Supplementary file Size Download File type/resource
GSM1071268_HX630-3.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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