|
| Status |
Public on Aug 28, 2018 |
| Title |
Mock_90min_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Murine L929 cells, mock-infected, 90min after actinomycin D treatment
|
| Organism |
Mus musculus |
| Characteristics |
cell line: fibroblast cell line L929
infection: mock-infected
agent: actinomycin D
time: 90min
|
| Treatment protocol |
L929 cells were plated in 150 x 25 mm tissue culture plates and were allowed to incubate at 37oC for 4 h, after which time the medium was removed and cells were mock-infected or infected with purified virions (Dearing, c87 or c8) at a multiplicity of infection of 80 plaque forming units/cell. After a 1.5 h viral adsorption at 37C, medium was added and samples were incubated at 37oC for an additional 18 h. At 19.5 h post-infection, actinomycin D was added at a final concentration of 10 µg/ml to stop transcription.
|
| Growth protocol |
Murine L929 cells were maintained as suspension cultures as described previously
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated at 0, 45, 90 and 120 min post-actinomycin D treatment using Trizol reagent according to the manufacturer’s instructions. Total RNA was purified with the RNeasy column.
|
| Label |
biotin
|
| Label protocol |
The purified cDNA was used for an in vitro transcription reaction using T7 RNA polymerase and biotinylated nucleotides following the manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Biotinylated anti-sense cRNA was purified with the RNeasy column; 15 µg was fragmented according to Affymetrix instructions’ and hybridized to Affymetrix murine U74Av2 oligonucleotide microarrays.
|
| Scan protocol |
Microarrays were scanned on a Hewlett Packard Agilent 2200 confocal scanner.
|
| Description |
Gene expression data from mock-infected cells 90min after adding actinomycin D
|
| Data processing |
Normalized signal intensities were obtained using Affymetrix MAS 5.0 software as described previously
|
| |
|
| Submission date |
Aug 27, 2018 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Liang Guo |
| Organization name |
University of Minnesota
|
| Street address |
2001 6th Street SE
|
| City |
minneapolis |
| State/province |
MINNESOTA |
| ZIP/Postal code |
55455 |
| Country |
USA |
| |
|
| Platform ID |
GPL81 |
| Series (1) |
| GSE119061 |
Reovirus infection induces stabilization and up-regulation of cellular transcripts that encode regulators of TGF-beta signaling |
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