Project description:Alveolar macrophages (AMs), the tissue dwelling monocytes of the lungs, are the first to encounter the menagerie of items that pass the mechanical barriers of the nose and throat. AMs must be able to mount an effective immune response against invading pathogens as well as maintain homeostasis in the lungs to avoid excessive inflammatory responses. In vivo data suggests that AMs can take on different phenotypes depending on the conditions of simulation. Limited progress has been made in defining the mechanisms responsible for these phenotypic issues largely due to logistical difficulties in isolating a sufficient number of cells from the lungs that have not been activated by the isolation protocol. To circumvent these logistical issues, an in vitro cell culture model was developed to study AM biology with specific emphasis on the generation of alternatively activated alveolar macrophages (AAAMs). Utilizing the BALB/c AM cell line MH-S it was determined that LPS could be used to generate classically activated AMs (CAAMs) and that IL-4 was effective in producing AAAMs. We examined the full transcriptome of AAAMs and CAAMs using microarray technology in an attempt to confirm previously described AAAM expression patterns and to identify new molecules associated with AM phenotypic change. AAAMs up-regulated an entirely distinct group of transcripts including genes encoding Arg1, Ym1 and a number of repair and remodeling genes whereas CAAMs up-regulated proinflammatory cytokines such as IL-1, IL-6 and IL-12. Additional novel markers are identified in this study to better characterize CAAMs and AAAMs in the lung. Keywords: Characterization of Cell Type Based on Activation Stimulus Overall design: MH-S Cells (ATCC CRL-2019) were stimulated with either 50 ng/ml IL-4 or 1µg/ml LPS and analyzed 12 hours later by whole genome DNA microarray analysis.
Project description:Comparing two subclones (Taiwan clone and Asian-Pacific clone) of CA-MRSA ST59. The Taiwan clone carries the Panton-Valentine leukocidin (PVL) genes, the staphylococcal chromosomal cassette mec (SCCmec) VT and is frequently isolated from patients with severe disease. The Asian-Pacific clone is PVL-negative, carries SCCmec IV, and is a frequent colonizer of healthy children.
Project description:We wished to identify Crl-regulated genes in stationary phase in E.coli, and whether those overlap with the previously identified regulon of RpoS. Therefore wildtype E.coli (MC4100) and its isogenic crl::cat mutant were grown at 30oC. Total RNA was extracted at on OD (578nm) of 4 (during entry into stationary phase) and then the analysis proceeded as described in detail in the protocoles. The experiment was repeated three times and the results from those experiments are presented here.
Project description:We identified a recurrent gene fusion TRA2B-DNAH5 in human lung squamous cell carcinoma. This gene fusion can promote tumor growth in CRL-5889 xenograft tumors. Microarary was performed to identify these differential genes which may mediate the tumor promotive function of TRA2B-DNAH5 fusion. Overall design: One human lung squamous carcinoma cell line CRL-5889 with or without TRA2B-DNAH5 fusion expression was used to do the xenograft assay in nude mice. 4 weeks later, we harvested the tumors and extract total RNAs for microarray analyses.
Project description:au13-04_cdtbis; cdt1_bis crl mutants and CDT1-RNAi lines have very similar macroscopic phenotypes as well as identical defects in plastid division and biogenesis. Our goal wwas to determine how much of these similarities originated from similar alterations of gene expression. Plantlets of crl mutant and CDT1-RNAi lines were grown in vitro on MS1/2 medium for 14 days. CDT1-RNAi lines were compared to the corresponding wild-type (Ws), whereas crl mutants were compared to their wild-type siblings and are in the Col0 ecotype. 4 dye-swap - normal vs rnai mutant comparaison, normal vs transgenic comparaison
Project description:Francisella tularensis LVS was grown in MH broth in the presence or absence of 200uM spermine. RNA was harvasted from overnight (16 hour) cultures and processed for microarray hybridization.
Project description:To investigate the dynamic regulation and co-regulation of miRNAs and mRNAs, a time-course experiment was performed as described before (E-MEXP-3544). Briefly, the time course experiment was carried out using the human A375 melanoma cell line (ATCC, CRL-1619).