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Sample GSM1126123 Query DataSets for GSM1126123
Status Public on Jul 14, 2014
Title Fibroblasts miR-132 mimic rep 2
Sample type RNA
Source name Fibroblasts transfected with miR-132 mimic
Organism Homo sapiens
Characteristics cell type: Human mammary GFP-tagged immortalized fibroblasts (218TGpp)
treatment: transfected with miR-132 mimic
Treatment protocol Fibroblasts were transfected with 25nM miRNA mimic (Dharmacon) using Lipofectamine (Invitrogen) in Opti-MEM medium depleted from serum and penicillin-streptomycin. The medium was changed to serum-containing medium 6h after transfection. After 24 hours, cells were either harvested for total RNA extraction (miRNeasy from Qiagen) or put in coculture.
Growth protocol Fibroblasts were grown in DMEM supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. Human mammary epithelial CCH cells were cultured in DMEM-F12 supplemented with 5% horse serum, cholera toxin (100 ng/ml), hydrocortisone (0.5 μg/ml), insulin (10 μg/ml), EGF (20 ng/ml) and 1% penicillin-streptomycin. All cells were maintained at 37C in a humified incubator (5% CO2).
Extracted molecule total RNA
Extraction protocol Transfection with miRNA mimics and total RNA extraction was perormed according to manufactures instructions.
Label biotin
Label protocol Standard Affymetrix protocol performed by AROS Applied Biotechnology A/S (Aarhus, Denmark). Biotinylated cDNA were prepared according to the standard Affymetrix protocol from 1 ug total RNA (GeneChip® Whole Transcript (WT) Sense Target Labeling Assay Manual, Version 4, Affymetrix).
Hybridization protocol Standard Affymetrix protocol performed by AROS Applied Biotechnology A/S (Aarhus, Denmark). Following fragmentation, cDNA was hybridized for 16 hr at 45C on GeneChip Human Gene 1.0 ST Arrays, followed by washing and staining using buffers from the GeneChip® Hybridization, Wash and Stain Kit.
Scan protocol Standard Affymetrix protocol performed by AROS Applied Biotechnology A/S (Aarhus, Denmark).
Description Gene expression from mammary fibroblasts transfected with miR-132 mimic
Data processing The data were analyzed with R, Bioconductor and Ingenuity Systems (Redwood City, CA). The RMA was used to normalise and summarise the expression values at the probeset level.
Submission date Apr 18, 2013
Last update date Jul 14, 2014
Contact name Sofie Björner
Organization name Lund University
Street address Jan Waldenströms gata 59
City Malmö
ZIP/Postal code 20502
Country Sweden
Platform ID GPL10739
Series (2)
GSE46197 Expression data from miR-132 overexpressing immortalized human mammary fibroblasts and from mammary epithelial columnar cell hyperplasia cells co-cultured with miR-132 overexpressing fibroblasts
GSE46199 Columnar cell hyperplasia

Data table header descriptions
VALUE log2 RMA signal

Data table
7892501 4.57106
7892502 5.26855
7892503 3.2675
7892504 9.13021
7892505 3.72496
7892506 6.07202
7892507 5.83167
7892508 6.89805
7892509 11.15913
7892510 4.08783
7892511 4.92425
7892512 7.69124
7892513 4.71112
7892514 10.46036
7892515 9.62305
7892516 5.17566
7892517 6.10892
7892518 2.76674
7892519 5.61976
7892520 8.98351

Total number of rows: 257430

Table truncated, full table size 4029 Kbytes.

Supplementary file Size Download File type/resource
GSM1126123_fibro_test_2.CEL.gz 4.1 Mb (ftp)(http) CEL
Processed data included within Sample table

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