|
| Status |
Public on Jan 08, 2011 |
| Title |
uninfected biological replicate 1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Caspase-1-/- bone marrow derived macrophages
|
| Organism |
Mus musculus |
| Characteristics |
treatment: uninfected
strain: B6
|
| Treatment protocol |
Macrophages were infected with the indicated bacterial strains at an MOI of 1. RNA was harvested 6h post transfection.
|
| Growth protocol |
Caspase-1-/- bone marrow derived macrophages on the B6 background were grown in RPMI + 10%FBS, 10%M-CSF, glutamine, and antibiotic.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNeasy Kit was used to extract total RNA.
|
| Label |
Cy5
|
| Label protocol |
RNA was amplified using the Ambion Amino Allyl MessageAmp Kit to incorporate amino-allyl-UTP. aRNA was labeled with Cy3 or Cy5 for 1h.
|
| |
|
| Channel 2 |
| Source name |
total RNA pooled from each experimental sample of mouse macrophages
|
| Organism |
Mus musculus |
| Characteristics |
reference: total RNA pooled from each experimental sample of mouse macrophages
strain: B6
|
| Treatment protocol |
Macrophages were infected with the indicated bacterial strains at an MOI of 1. RNA was harvested 6h post transfection.
|
| Growth protocol |
Caspase-1-/- bone marrow derived macrophages on the B6 background were grown in RPMI + 10%FBS, 10%M-CSF, glutamine, and antibiotic.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNeasy Kit was used to extract total RNA.
|
| Label |
Cy3
|
| Label protocol |
RNA was amplified using the Ambion Amino Allyl MessageAmp Kit to incorporate amino-allyl-UTP. aRNA was labeled with Cy3 or Cy5 for 1h.
|
| |
|
| |
| Hybridization protocol |
Labeled aRNA probe was hybridized in 3X SSC, 25mM HEPES pH 7, 0.25%SDS for 36h at 63°C. Slides were washed and dried before scanning.
|
| Scan protocol |
Scanned on a GenePix 2000 Scanner.
|
| Data processing |
Background was subtracted, data were normalized linearly by ratio of medians, and the data are presented as log2 of Cy5/Cy3. Acuity software was used.
|
| |
|
| Submission date |
Jan 06, 2011 |
| Last update date |
Jan 08, 2011 |
| Contact name |
Russell Vance |
| E-mail(s) |
rvance@berkeley.edu
|
| Phone |
510-643-2795
|
| Organization name |
UC Berkeley
|
| Street address |
415 Life Science Addition
|
| City |
Berkeley |
| State/province |
CA |
| ZIP/Postal code |
94706 |
| Country |
USA |
| |
|
| Platform ID |
GPL5137 |
| Series (2) |
| GSE26473 |
Secreted bacterial effectors that inhibit host protein synthesis are critical for induction of the innate immune response to virulent Legionella pneumophila [exp1] |
| GSE26491 |
Secreted bacterial effectors that inhibit host protein synthesis are critical for induction of the innate immune response to virulent Legionella pneumophila |
|
