|
| Status |
Public on Sep 23, 2015 |
| Title |
NTHi375 infected A549 cells_2hr_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
NTHi375 infected A549 cells_2hr_rep1
|
| Organism |
Homo sapiens |
| Characteristics |
agent: Haemophilus influenzae strain NTHi375
cell line: A549
|
| Treatment protocol |
A freshly grown A549 cell monolayer was infected in EBSS medium with Haemophilus influenzae strain NTHi375, using a multiplicity of infection of 100:1, for 2 h at 37 ºC in a humidified incubator with 5% CO2. As a control, a freshly grown A549 cell monolayer was left untreated.
|
| Growth protocol |
A549 cells were grown for 32 h with RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum at 37ºC in a humidified incubator with 5% CO2 to reach 90% confluency; at this point, cells were serum starved for 16 h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using a Nucleospin RNAII kit (Macherey-Nagel) as recommended by the manufacturer, including an on column DNase treatment step. Total RNA quality was evaluated using RNA 6000 Nano LabChips (Agilent 2100 Bioanalyzer, Santa Clara, CA). All samples had intact 18S and 28S ribosomal RNA bands with RNA integrity numbers (RIN) between 9.3 to 9.7, and RNA A260/280 ratios of 2.1.
|
| Label |
Cy3
|
| Label protocol |
Samples Labeling was performed using the “Amino Allyl MessageAmp II aRNA Amplification Kit (Ambion)” following manufacturer recommendations. Each sample was fluorescently labeled with Cy3-fluorescent dye.
|
| |
|
| Hybridization protocol |
Cy3 fluorescently labeled samples were hybridized to “Agilent (012097) Human 1A Microarrays” according to manufacturer recommendations (Agilent Technologies, Santa Clara, CA, USA) recommendations.
|
| Scan protocol |
Slides were scanned using a Gene Pix 4100A scanner (Axon Instruments, Union City, CA, USA) at 5µ resolution. Image analysis was performed using associated software GenePiX Pro.
|
| Description |
Gene expression analysis of A549 human type II pneumocyte infected with Haemophilus influenzae strain NTHi375 for 2 h post-infection.
|
| Data processing |
The data were normalized with Bioconductor software using quantiles.
|
| |
|
| Submission date |
May 21, 2015 |
| Last update date |
Sep 23, 2015 |
| Contact name |
Junkal Garmendia |
| E-mail(s) |
juncal.garmendia@unavarra.es
|
| Organization name |
Instituto de Agrobiotecnología-CSIC-UPNA-Gob
|
| Department |
Sanidad Animal
|
| Street address |
Avda de Pamplona, 123
|
| City |
Pamplona |
| ZIP/Postal code |
31193 |
| Country |
Spain |
| |
|
| Platform ID |
GPL887 |
| Series (1) |
| GSE69134 |
Genome expression profiling identifies host-directed antimicrobial drugs against respiratory infection by nontypable Haemophilus influenzae |
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