Project description:Total gene expression analysis was performed on constitutive H3f3b KO E12.5 MEFs relative to WT littermates. Intent was to analyze the role of H3f3b in overall gene expression. Constitutive KO mice were obtained by mating H3f3b Fl/Fl mice to Zp3Cre mice. Heterozygous offspring were time mated and embryos were isolated 12.5 days after. MEFs were cultured transduced with a stable pBabe puro vector and total RNA was isolated from H3f3b KO and WT MEFs.
Project description:Total gene expression analysis was performed on RNA from testes extracted from two litters of constitutive homozygous and heterozygous H3f3b knockout mice compared to WT littermates. Constitutive knockout mice were obtained by mating H3f3b Fl/Fl mice to Zp3Cre mice. Heterozygous knockout mice were crossed until homozygous knockout mice were obtained. Testes were surgically removed from 8 week old homozygous knockouts, heterozygous knockouts and WT, and RNA was extracted from one testis from each mouse for total gene expression.
Project description:Total gene expression analysis was performed on CRE induced conditional knockout E12.5 MEFs relative to GFP infected control MEFs. Intent was to analyze the role of H3f3b in overall gene expression. For conditional KO, three lines of H3f3b FL/FL E12.5 MEFs (cKO1, cKO2 and pcKO1) were transduced with Cre retroviruses and compared to their respective lines of MEFs transduced solely with GFP vector (as control). Total RNA was isolated for gene analysis.
Project description:A mixed-aerosol pH1N1 (Cal04) challenge of rhesus macaques was establised to serve as a pre-clinical model for the evaluation of candidate vaccines. After characterizing the clinical signs and immune responses associated with pH1N1 challenge in naïve rhesus macaques, a follow-up study assessing 2 candidate vaccines was performed. This study has 2 phases: 1) Model Establisment consisting of 3 groups: Unvaccinated Live Challenge (n=3, Unvaccinated UV-inactivated Challenge (n=3), Previously Vaccinated Live Challenge (n=3) which were sampled at 2 baseline timepoints Day -7 and Day 0. Following the H1N1 Challenge, samples were collected at day 1,2,5,8,14,20. 2) Candidate Vaccine Assessment consisting of four groups: Previously Vaccinted with anti-CD40-NP5+PolyICLC (n=4), Previously vaccinated with CD40-HA+PolyICLC (n=4), Previously vaccinated with commercial mismatched Fluzone (n=4), Previously vaccinated with Media+PolyICLC alone (n=4). Daseline samples were collected at Day -7 and 0 (baseline) and Day 1,3,6,14,20 post-challenge.
Project description:The goal of this study is to identify global changes in gene expression that accompany induction of site-specific RNA editing that targets the SDHB gene in monocyte-enriched peripheral blood mononuclear cell cultures. The results provide important information to understand the function of this programmed SDHB RNA mutation within the context of global gene expression changes that occur in cultured immune cells when the editing is induced. The study also specifically tests whether expression changes occur in SDH subunit genes, cytidine deaminase family of genes and hypoxia-inducible pathways. Overall design includes pairwise comparison of total gene expression profiles from uncultured cold-aggregated PBMCs (day 0), low-editing (culture day 3) and high-editing (culture days 5-8) samples, each from 4 donors. Submitted manuscript reported summary results from comparison of cultured low- and high-editing samples.
Project description:The DALIA-1 trial was designed to evaluate the safety and immunogenicity of a DC-based vaccine generated by culturing blood monocytes with GM-CSF and IFN-a. DCs were pulsed for 24 hrs with the ANRS LIPO5 peptide pool and activated for the last 6 hrs with LPS. In total, 19 asymptomatic HAART-treated patients were included in the study. Microarray samples were collected in both the treatment phase and follow-up phase of the study. Whole blood RNA samples were collected at two time points prior to the first vaccination (Weeks -8 and -4) and served as a pre-vaccination baseline. Additional whole blood RNA samples were collected at the time of vaccination (Weeks 0, 4, 8, 12) and post-vaccination at weeks 16, 22, and 24. At week 24, an analytical treatment interruption (ATI) was initiated during which HAART was suspended. During this post-ATI follow-up phase, whole blood RNA samples were collected at weeks 25, 26, 27, 28, 32, 36, 40, , 44, and 48). Additionally, at weeks -4, 16, and 48 patients were apheresed. 18 samples are constituents of both the pre-ATI and post-ATI datasets, which were normalized separately. The normalised data underwent the following preprocessing: First a normal-exponential convolution model was applied (Xie et al, Bioinformatics, 2009; Shi et al, Nucleid Acids Research, 2010) to correct for the background noise. An offset of 16 (default value) was then added to the data (to improve precision and avoid a compression of the fold changes) before a quantile-quantile normalization was performed. Finally, data were log2 transformed. Only probes whose pval-detection (provided by Illumina Genome-Studio software) was below 0.01 in at least one sample were kept further on. At last, the ComBat method (Johnson et al, Biostatistics, 2007) was applied to those data in order to correct for batch effects (namely a small hybridization chamber effect, that was identified through Principal Variance Component Analysis - Li et al, chap. 12, Batch Effects and Noise in Microarray Experiments: Sources and Solutions, Wiley, 2009).
Project description:This SuperSeries is composed of the following subset Series: GSE40560: Transcriptome analysis in primary fibroblasts from HOIL-1-deficient patients upon TNF-alpha or IL-1beta stimulation GSE40561: Transcriptional analysis of whole blood in patients with auto-inflammatory disorders GSE40838: Transcriptome analysis in peripheral blood mononuclear cells (PBMC) from HOIL-1-deficient patients upon TNF-alpha or IL-1beta stimulation Refer to individual Series
Project description:Differential gene expression study comparing choriogonadotropin alpha (Cga) gene deficient and wild type adult mouse pituitary. Thyrotrope hyperplasia and hypertrophy are common responses to primary hypothyroidism. To understand the genetic regulation of these processes, we studied gene expression changes in the pituitaries of Cga-/- mice, which are deficient in the common alpha subunit of TSH, LH and FSH. These mice have thyrotrope hypertrophy and hyperplasia and develop thyrotrope adenoma. We report that cell proliferation is increased, but the expression of most stem cell markers is unchanged. The alpha subunit is required for secretion of the glycoprotein hormone beta subunits, and mutants exhibit elevated expression of many genes involved in the unfolded protein response, consistent with dilation and stress of the endoplasmic reticulum. Mutants have elevated expression of transcription factors that are important in thyrotrope function, such as Gata2 and Isl1, and those that stimulate proliferation, including Nupr1, E2f1 and Etv5. We characterized the expression and function of a novel, over-expressed gene, transcription elongation factor A (SII)-like 5 (Tceal5). Stable expression of Tceal5 in a pituitary progenitor cell line is sufficient to increase cell proliferation. Thus, Tceal5 may act as a proto-oncogene. This study provides a rich resource for comparing pituitary transcriptomes and an analysis of gene expression networks. Total RNA was obtained from 8 week old Cga-deficient and littermate wild type mouse pituitary tissue. Total sample number was 24 as follows: 6 male and 6 female wild types, 6 male and 6 female mutants. Each sample was applied to an individual microarray.