Dataset Information


Murine lung transcripts during soluble egg antigen (SEA)-induced allergic asthma and bleomycin-induced pulmonary fibrosis in Wildtype, IL-13Ra1 and IL-13Ra2 deficient mice sensitized and challenged

ABSTRACT: Transcriptional profiling of mouse lungs by comparing PBS and saline treated lungs with SEA and bleomycin treated lungs, respectively. Mice strains used in this study include wild type (C57BL/6), IL-13Ra1 and IL-13Ra2 deficient mice. Overall design: In mouse model of allergic asthma, mice were sensitized twice (day0 and day14) by i.p. injection of 10 µg of SEA. On days 28 and 31 mice were anesthetized with a mixture of xylazine and ketamine and given an intratracheal (I.T) airway challenge with 10 µg of SEA. Mice were sacrificed 24 h after the final airway challenge (day 32) and lungs were collected for RNA preparation. Biological replicates include 5 PBS treated and 5 SEA treated mice. In bleomycin-induced fibrosis odel, mice were anaesthetized with a xylazine and ketamine cocktail and given 0.15 U bleomycin sulfate (EMD) in saline or saline alone via the I.T. route. On day 7, mice were sacrificed and ungs were collected for RNA preparation. Biological replicates include 4 saline treated and 4 bleomycin treated mice. Goal was to determine IL-13 receptors regulated genes during SEA induced allergic asthma and bleomycin-induced fibrosis. Fluorescent cDNA targets were prepared from a 20 μg experimental RNA sample (SEA or bleomycin challenged group–dUTPCy5 – Amersham, Piscataway, NJ) and a 20 μg reference RNA sample (PBS or saline treated group–dUTPCy3- Amersham, Piscataway, NJ). Equal quantities of the above labeled cDNA (experimental and reference labeled RNA samples) were mixed and any free label present in the sample was removed by washing 3 times using a 10 kDa cutoff Vivaspin filters (Millipore). Labeled fluorescent cDNA targets were hybridized on the Whole Mouse Genome Oligo Microarray Kit (Agilent, Palo Alto, CA) containing more than 41000 gene probes.

INSTRUMENT(S): NIAID Mmeb -- Mouse 44k - mAdb version

ORGANISM(S): Mus musculus  

SUBMITTER: Sudhir Varma  

PROVIDER: GSE27721 | GEO | 2011-09-07



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