Characterization of Lung MDSC from SKG mice

The purpose of this project is to characterize the myeloid derived suppressor cells (MDSCs) in the inflamed lung of interstitial lung disease SKG mice. We compared the gene expression of lung MDSCs with MDSCs from bone marrow or spleen.BD FACSAria III (BD Biosciences) was used to sort CD11b+Gr1+ cells (MDSCs) from BM, spleen and lung of interstitial lung disease model SKG mice. Total RNA was extracted from cultured cells using Maxwell RSC simplyRNA Cells Kit (Promega). Biotinylated cRNA were prepared using GeneChip 3'IVT PLUS Reagent Kit (Thermo Fisher Scientific) according to the standard manufacturer's protocol from 100 ng total RNA. Following fragmentation, 12.5 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Mouse Genome 430 2.0 Array. GeneChips were washed and stained in the GeneChip Fluidics Station 450. GeneChips were scanned using the GeneChip Scanner 3000 7G. The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Expression Console Software (ThermoFisher Scientific) according to the default analysis settings.