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Sample GSM420245 Query DataSets for GSM420245
Status Public on Nov 19, 2009
Title SH-SY5Ycells_bilirubin_treated_140nM_1hour_rep2
Sample type RNA
 
Source name Human neuroblastoma SH-SY5Y cells, treated with free bilirubin (Bf) 140nM, for 1 hour
Organism Homo sapiens
Characteristics cell line: SH-SY5Y
atcc: CRL-2266
cell type: neuroblastoma
treatment: free bilirubin
treatment time: 1 hour
Treatment protocol The growth medium was replaced with experimental medium, which was either 140 nM Bf medium (Bf 140 nM samples) or 0.6 % DMSO medium (DMSO 0.6 % samples), and cells were allowed to grow. After 1 h, cells were collected and processed for microarray analysis. Separate flasks of cells were used for each of the treatments and assays. Experiments were performed using at least three independent biological replicates.
Growth protocol SH-SY5Y human neuroblastoma cells (ATCC CRL-2266) were cultured in a humidified incubator at 37°C and 5 % CO2 in F12-EMEM (1:1) containing 15 % foetal bovine serum (FBS), 2 mM L-glutamine, 25 µg/mL gentamicin, 100 U/ml penicillin, and 100 µg/mL streptomycin at 37°C with 5 % CO2. Cells were seeded at 6 x 10e6 cells in 75 cm2 flasks and grown for 24 h (cells reached 70-80% confluence).
Extracted molecule total RNA
Extraction protocol Total RNA was isolated using the TRIzol reagent (Invitrogen) following the manufacturer’s instructions. It was then purified using the RNeasy mini kit (Qiagen). The quality of total RNA was assessed using a bioanalyzer (Agilent 2100; Agilent Technologies) and RNA was quantified by using a ND-1000 Nanodrop spectrophotometer.
Label Biotin
Label protocol 10 μg of each total RNA sample was labelled according to the standard one-cycle amplification and labelling protocol developed by Affymetrix (Santa Clara, CA).
 
Hybridization protocol Labelled cRNA was hybridized on Affymetrix GeneChip Human U133A 2.0 Arrays containing over 14,500 transcripts. Hybridized GeneChips were stained and washed using the GeneChip Fluidic Station 450.
Scan protocol GeneChips were scanned with the GeneChip Scanner 3000 7G. Cell intensity values and probe detection calls were computed from the raw array data using the Affymetrix GeneChip Operating Software (GCOS).
Description Bf140nM1h_5
Data processing Further data processing was performed in the R computing environment (http://www.r-project.org/, version R 2.5.0 for Windows) using packages from the BioConductor software project (http://www.bioconductor.org/). Variance-stabilizing normalization was applied, using the justvsn function from the vsn library. Normalized data were then filtered based on the Affymetrix detection call, so that only probes that had a Present call in at least one of the arrays were retained. Data were then imported in the MultiExperiment Viewer (MeV) software (version 4.0.01 for Windows XP), and statistical analysis was performed with the SAM (Significance Analysis of Microarrays) module.
 
Submission date Jun 23, 2009
Last update date Nov 19, 2009
Contact name Paola Roncaglia
Organization name SISSA/ISAS (International School for Advanced Studies)
Department Neurobiology
Street address via Bonomea 265
City Trieste
State/province TS
ZIP/Postal code 34136
Country Italy
 
Platform ID GPL571
Series (2)
GSE16766 Transcriptome analysis identifies molecular effectors of unconjugated bilirubin in human neuroblastoma SH-SY5Y cells: 1h
GSE16768 Transcriptome analysis identifies molecular effectors of unconjugated bilirubin in human neuroblastoma SH-SY5Y cells

Data table header descriptions
ID_REF
VALUE VSN signal

Data table
ID_REF VALUE
1007_s_at 9.834345355
1053_at 10.34606936
117_at 6.958794115
121_at 8.876232505
1255_g_at 8.086684314
1294_at 7.290596609
1316_at 7.099362748
1320_at 7.721737627
1405_i_at 6.034108507
1431_at 6.249071146
1438_at 7.852518934
1487_at 8.958202886
1494_f_at 7.657253961
1598_g_at 9.591053234
160020_at 7.73266641
1729_at 8.628638316
177_at 6.580378644
1773_at 8.413787579
179_at 9.551236113
1861_at 8.65026283

Total number of rows: 22277

Table truncated, full table size 496 Kbytes.




Supplementary file Size Download File type/resource
GSM420245.CEL.gz 2.1 Mb (ftp)(http) CEL
Processed data included within Sample table

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