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Sample GSM1179371 Query DataSets for GSM1179371
Status Public on Jul 03, 2013
Title wdNHBE with KY/180 at 36hpi, biological rep2
Sample type RNA
 
Source name Well-differentiated human lung epithelial cells, infected with KY/180 (pdmH1N1) for 36h
Organism Homo sapiens
Characteristics infection: 2009 pandemic H1N1 Influenza virus
cell type: cultured primary human lung epithelial cells
Treatment protocol Infection of wd-NHBE cells were conducted in serum-free growth media provided by manufacturer (MatTek). Wd-NHBE cells, cultured at the air-liquid interface, were washed two times with Dulbecco’s phosphate buffered saline (DPBS) to remove mucus accumulation. Virus stocks were diluted in DPBS and cells were infected at a multiplicity of infection (MOI) of 3.0 for 1 h at 37°C, 5% CO2. Cells were washed twice with DPBS to remove unbound virus. Basal medium was removed and replaced with complete medium. Over the time course of infection, the basal media was removed and apical layer washed twice with DPBS supplemented with 0.2% BSA (Sigma) and stored at -80 degrees celcius until use.
Growth protocol well-differentiated cells (wd-NHBE) were purchased from MatTek Corporation (EpiAirway PC-12, Ashland, MA). Cells were maintained by incubating the 12-well formatted transwell inserts at the air-liquid interface at 37°C under 5% CO2. The basal surface was submerged in AIR 100 (MatTek) complete growth media, made up of DMEM , epidermal growth factors, 5 µg/ml gentamicin, 0.25 µg/ml amphotericin B, and phenol red, and was replaced every 24 h until used for experiments.
Extracted molecule total RNA
Extraction protocol Trizol extraction of total RNA was performed according to the manufacturer's instructions.
Label biotin
Label protocol Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 100ng total RNA (Gene Chip 3' IVT Expression Kit User Manual, 2008, Affymetrix).
 
Hybridization protocol Following fragmentation, 12.5ug of aRNA were hybridized for 16 hr at 45C on GeneChip Human (HG_U133 plus 2.0) Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450/250.
Scan protocol Scanning of chip using Affymetrix Command Console (version 3.1)
Description Gene expression data from well-differentiated primary human lung bronchial epithelial cells infected with H1N1
Data processing Prior to statistical analyses, raw data were processed by Plier Workflow normalization method using Gene Console software (Affymetrix, version 1.3.1).
 
Submission date Jul 02, 2013
Last update date Jul 03, 2013
Contact name Rachael Gerlach
E-mail(s) rmlask01@louisville.edu
Organization name University of Louisville
Department Microbiology and Immunology
Lab Jonsson
Street address 505 S Hancock, CTR 626
City Louisville
State/province KY
ZIP/Postal code 40202
Country USA
 
Platform ID GPL570
Series (1)
GSE48466 Expression data from well-differentiated human bronchial epithelial cells infected with H1N1 Influenza isolates

Data table header descriptions
ID_REF
VALUE Normalized data

Data table
ID_REF VALUE
AFFX-BioB-5_at 200.1455
AFFX-BioB-M_at 302.2263
AFFX-BioB-3_at 220.6029
AFFX-BioC-5_at 509.451
AFFX-BioC-3_at 662.2336
AFFX-BioDn-5_at 1567.771
AFFX-BioDn-3_at 2288.352
AFFX-CreX-5_at 7201.409
AFFX-CreX-3_at 7303.651
AFFX-DapX-5_at 147.0202
AFFX-DapX-M_at 436.5578
AFFX-DapX-3_at 615.4884
AFFX-LysX-5_at 33.46108
AFFX-LysX-M_at 45.0239
AFFX-LysX-3_at 84.78342
AFFX-PheX-5_at 58.19198
AFFX-PheX-M_at 73.56355
AFFX-PheX-3_at 70.27148
AFFX-ThrX-5_at 53.65841
AFFX-ThrX-M_at 97.04472

Total number of rows: 54675

Table truncated, full table size 1054 Kbytes.




Supplementary file Size Download File type/resource
GSM1179371_KY180_2_36hpi.CEL.gz 4.8 Mb (ftp)(http) CEL
Processed data included within Sample table

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