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Sample GSM3018255 Query DataSets for GSM3018255
Status Public on Feb 21, 2018
Title HEC-TM-OHT24h_R3
Sample type RNA
Source name Mammary epithelial cells immortalized with hTERT treated with 4-OHT during 24 hours
Organism Homo sapiens
Characteristics cell type: Human mammary epithelial cells (HECs; Lonza)
treatment: treated with 4-OHT during 24 hours
Treatment protocol HECs were treated daily for 24 or 96 hours with 4-OHT (Sigma-Aldrich) at 250 nM, or with 65 mM KCl (Sigma-Aldrich),for 24 hours
Growth protocol Human mammary epithelial cells (HECs; Lonza) were cultured in mammary epithelial cell growth medium (Promocell) with penicillin/ streptomycin 100 U/ml (Life Technologies). The cells were maintained in a humidified atmosphere at 37°C under 5% CO2.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted from HEC-TM cells using NucleoSpin® RNA (Macherey Nalgen) following the manufacturer's recommendations
Label Cy3
Label protocol cRNAs were synthesized and labeled with the Cy3 dye from 100 ng of total RNA using the one-color Low Input Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer's recommandations, followed by RNeasy Mini column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
Hybridization protocol 1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. After fragmentation 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent GE Whole Human Genome Oligo Microarrays (Agilent-026652) 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 briefly in acetonitrile.
Scan protocol Microarrays were scanned with an Agilent DNA microarray scanner G2565CA (Agilent Technologies).Fluorescent signals were extracted and normalized with the Feature Extraction software version (Agilent Technologies),
Data processing The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
Submission date Feb 20, 2018
Last update date Feb 21, 2018
Contact name jean-michel flaman
Phone +33687477812
Organization name Inserm 1052
Department CRCL
Street address 28 rue Laennec
City Lyon
ZIP/Postal code 69373
Country France
Platform ID GPL13497
Series (1)
GSE110884 The SCN9A channel and plasma membrane depolarization promote cellular senescence through Rb pathway

Data table header descriptions
VALUE Data were normalized to the 75th percentile signal intensity and the baseline was adjusted on control HEC-TM non treated condition using GeneSpring GX 12.6 software (Agilent Technologies)

Data table
GE_BrightCorner -0.3462677
DarkCorner -0.9181099
A_23_P146146 -0.89245796
A_23_P42935 0.2649498
A_23_P117082 0.08168316
A_23_P2683 0.16228867
A_24_P358131 -0.059627533
A_33_P3367647 -0.881382
A_23_P157316 -0.119125366
A_32_P14850 -0.108716965
A_23_P158596 -0.52809143
A_23_P350107 -0.16664314
A_23_P388190 -0.032780647
A_23_P106544 0.058927536
A_33_P3219745 -0.8671093
A_32_P85539 -0.08988857
A_23_P94998 0.04298687
A_33_P3235677 -0.8613243
A_23_P417014 -0.58707285
A_23_P103905 -0.008790016

Total number of rows: 34183

Table truncated, full table size 820 Kbytes.

Supplementary file Size Download File type/resource
GSM3018255_HEC-TM-OHT24h_R3.txt.gz 2.1 Mb (ftp)(http) TXT
Processed data included within Sample table

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