Transcriptome Characterization of Bovine Peripheral Blood Mononuclear Cells Stimulated by LPS
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ABSTRACT: This study aimed to identify transcription networks and signatures of bovine peripheral blood mononuclear cells in response to LPS stimulation. A total of 464 genes including at least 17 transcription factors were identified to be significantly induced by LPS using a high density bovine oligonucleotide microarray. The network analysis revealed that alternation in gene expression was regulated by transcription factors through potential interaction within the pathway networks in the LPS stimulated cells. Our results demonstrated that specific pathway networks are responsible for transcription regulation in bovine peripheral blood mononuclear cells in response to pathogen components such as LPS. One class unpaired: Bovine Peripheral Blood Mononuclear Cells from 10 BLV positive cattle are divided into 2 groups: 5 treated with PBS in vitro for 2h serve as controls. The remaining 5 samples are stimulated with LPS in vitro for 2h.
Project description:Background: Genome-wide detection of single feature polymorphisms (SFP) in swine using transcriptome profiling of day 25 placental RNA by contrasting probe intensities from either Meishan or an occidental composite breed with Affymetrix porcine microarrays is presented. A linear mixed model analysis was used to identify significant breed-by-probe interactions. Results: Gene specific linear mixed models were fit to each of the log2 transformed probe intensities on these arrays, using fixed effects for breed, probe, breed-by-probe interaction, and a random effect for array. After surveying the day 25 placental transcriptome, 789 probes with a q-value ⤠0.05 and |fold change| ⥠2 for the breed-by-probe interaction were identified as candidates containing SFP. To address the quality of the bioinformatics approach, universal pyrosequencing assays were designed from Affymetrix exemplar sequences to independently assess polymorphisms within a subset of probes. Of those probes sampled from high-, medium-, and low-ranking categories, 20 of 27 were confirmed by pyrosequencing to contain SFPs. In most cases, the 25-mer probe sequence printed on the microarray diverged from Meishan, not occidental crosses. This analysis was used to define a set of highly reliable predicted SFPs according to their probability scores. Conclusions: By this method we detected transition and transversion single nucleotide polymorphisms, as well as insertions/deletions. These results demonstrate that this approach can identify polymorphisms between two breeds and/or lines of any species for which a short oligonucleotide array is available, and can be used to rapidly develop markers for genetic mapping and association analysis in species where high density genotyping platforms are otherwise unavailable. SNPs and INDELS discovered by this approach have been publicly deposited in NCBIâs SNP repository dbSNP. This method is an attractive bioinformatics tool for uncovering breed-by-probe interactions, for rapidly identifying expressed SNPs, for investigating potential functional correlations between gene expression and breed polymorphisms, and is robust enough to be used on any Affymetrix gene expression platform. Keywords: Transcriptional profiling of Day 25 porcine placentas 6 samples: Breed (Meishan, Occidental) Placental Tissue, Day 25
Project description:Using two complementary approaches, analysis of imprinting of candidate genes by pyrosequencing and expression profiling of parthenogenetic fetuses, we carried a comprehensive survey of genomic imprinting in swine. In the case of imprinted genes where transcription of one of the two parental alleles is silenced, uniparental embryos like parthenotes can be used to measure transcript dosage effects. Using Affymetrix Porcine GeneChip microarrays, four tissues of day 30 fetuses were profiled: brain, fibroblast, liver, and placenta. Keywords: Transcriptional profiling of epigenetic asymmetry in porcine day 30 parthenogenetic and biparental fetal tissues 24 samples total of Day 30 occidental porcine fetuses: 2 treatments (parthenote, control); four tissues (brain, fibroblast, liver, placenta); technical replicates (3 fibroblast only)
Project description:Background: Genome-wide detection of single feature polymorphisms (SFP) in swine using transcriptome profiling of day 25 placental RNA by contrasting probe intensities from either Meishan or an occidental composite breed with Affymetrix porcine microarrays is presented. A linear mixed model analysis was used to identify significant breed-by-probe interactions. This dataset uses different tissue types to explore the consequences of additional sample variance. Keywords: Transcriptional profiling of porcine tissues 6 samples: 3 Day 30 occidental fibroblast, 3 Day 25 Meishan placenta.
Project description:Tiled regions surrounding 5 human genes as 36mers, HBG2, TIMP3, SYN3, FLNA, FBX07. The first three of these genes, we tiled with various mismatch oligos in addition to 'perfect match' oligos. Keywords: Mismatch hybridization experiment Tiled perfect match and various designs of mismatch oligonucleotide for several human genes. Goal was to observe the influence of various MM types on hybridization behavior in human, and compare it to yeast (see related slide).
Project description:A unifying characteristic of aggressive cancers is a profound anabolic shift in metabolism to enable sustained proliferation and biomass expansion. The ribosome is centrally situated to sense metabolic states but whether it impacts systems that promote cellular survival is unknown. Here, through integrated chemical-genetic analyses, we find that a dominant transcriptional effect of blocking protein translation in cancer cells is complete inactivation of heat shock factor 1 (HSF1), a multifaceted transcriptional regulator of the heat-shock response and many other cellular processes essential for tumorigenesis. Translational flux through the ribosome reshapes the transcriptional landscape and links the fundamental anabolic processes of protein production and energy metabolism with HSF1 activity. Targeting this link deprives cancer cells of their energy and chaperone armamentarium thereby rendering the malignant phenotype unsustainable. Gene Expression Data We used microarrays to examine affect of rocaglates on gene expression in cancer cell lines. MCF7 breast cancer cells were treated with either 200 nM Rocaglamide A or DMSO, as a control. Two biological replicates for all samples.
Project description:Sexual dimorphism in mammals is mostly attributable to sex-related hormonal differences in fetal and adult tissues; however, this may not be the sole determinant. Though genetically-identical for autosomal chromosomes, male and female preimplantation embryos could display sex-specific transcriptional regulation which can only be attributted to the differences in sexual chromosome dosage. We used microarrays to analyze sex-related transcriptional differences at the blastocyst stage. Day 7 bovine in vitro produced bovine blastocysts produced with sorted semen from 3 different bulls. Pooled RNA from 60 blastocysts of one sex and produced with one bull was used per chip. Three replicates of each sex per bull. In total, 18 Bovine GeneChip (Affymetrix) were used (3 replicates X 3 bulls X 2 sexes).
Project description:Heat-Shock Factor 1 (HSF1), master regulator of the heat-shock response, facilitates malignant transformation, cancer cell survival and proliferation in model systems. The common assumption is that these effects are mediated through regulation of heat-shock protein (HSP) expression. However, the transcriptional network that HSF1 coordinates directly in malignancy and its relationship to the heat-shock response have never been defined. By comparing cells with high and low malignant potential alongside their non-transformed counterparts, we identify an HSF1-regulated transcriptional program specific to highly malignant cells and distinct from heat shock. Cancer-specific genes in this program support oncogenic processes: cell-cycle regulation, signaling, metabolism, adhesion and translation. HSP genes are integral to this program, however, even these genes are uniquely regulated in malignancy. This HSF1 cancer program is active in breast, colon and lung tumors isolated directly from human patients and is strongly associated with metastasis and death. Thus, HSF1 rewires the transcriptome in tumorigenesis, with prognostic and therapeutic implications. We used microarrays to examine affect of HSF1 depletion on gene expression in cancer cell lines. Three cancer cell lines (BPLER, HMLER & MCF7) were transduced with either control shRNAi (Scramble or GFP) or an shRNAi that targets and depletes HSF1 (hA6). Another BPLER sample was subjected to a 1H, 42M-KM-^Z C heat shock. Two biological replicates for all samples.
Project description:This study aimed to identify transcription networks and signatures of bovine peripheral blood mononuclear cells in response to LPS stimulation. A total of 464 genes including at least 17 transcription factors were identified to be significantly induced by LPS using a high density bovine oligonucleotide microarray. The network analysis revealed that alternation in gene expression was regulated by transcription factors through potential interaction within the pathway networks in the LPS stimulated cells. Our results demonstrated that specific pathway networks are responsible for transcription regulation in bovine peripheral blood mononuclear cells in response to pathogen components such as LPS.
Project description:Rgg-dependent transcriptional regulation in Streptococcus pyogenes strains MGAS5005 and CS101 was analyzed during post-exponential phase of growth Keywords: strain comparion, post-exponential growth, rgg mutant Microarray analysis was performed using RNA samples isolated from wild-type MGAS5005 and CS101 strains as well as their rgg mutant strains during post-exponential phase of growth