Difference in RNA content of different cell types introduces bias to gene expression deconvolution methods. If ERCC spike-ins are introduced into samples, predicted proportions of deconvolution methods can be corrected Overall design: Two cell types of distinctly different sizes and RNA per cell content: HEK cells and Jurkat cells were mixed in different proportions ensuring that each mixture contained total of one million cells. We sequenced RNA of the samples (including ERCC spike-in controls to 382 be able to control for the absolute RNA-concentration).
Complete deconvolution of cellular mixtures based on linearity of transcriptional signatures.
Cell line, Subject
View SamplesThe CMVpp65 protein contains 2 bipartite nuclear localization signals (NLS) at 415-438aa and 537-561aa near the carboxy terminus of CMVpp65 and a phosphate binding site related to kinase activity at lysine-436. A mutation of pp65 having K436N (CMVpp65mII) and further deletion of aa537-561 resulted in a novel protein (pp65mIINLSKO) that is kinase-less and has markedly reduced nuclear localization. The purpose of this report was to study the biologic characterization of this protein and its immunogenicity compared to native pp65.Using RNA microarray analysis, expression of the CMVpp65mIINLSKO had less effect on cell cycle pathways than did the native CMVpp65 and a greater effect on cell surface signalling pathways involving immune activity. It is concluded that the removal of the primary NLS motif from pp65 does not impair its immunogenicity and may actually be advantageous in the design of a vaccine.
Biologic and immunologic effects of knockout of human cytomegalovirus pp65 nuclear localization signal.
No sample metadata fields
View SamplesGranulomas are immune cell aggregates formed in response to persistent inflammatory stimuli. Granuloma macrophage subsets are diverse and carry varying copy numbers of their genomic information. The molecular programs that control the differentiation of such macrophage populations in response to a chronic stimulus, though critical for disease outcome, have not been defined. In this study, we performed scRNA-Seq experiments to gain insights into the transcriptional regulation of polyploid macrophage differentiation in response to chronically persistent inflammatory stimuli. Overall design: scRNA-Seq was performed on FACS-sorted 2c and >4c DNA content polyploid macrophages after six days of bacterial lipoprotein, FSL-1 treatment of bone marrow-derived macrophage precursors. 2c DNA content macrophages treated with M-CSF alone were used as controls. CEL-Seq2 protocol was used for single cell sequencing (Hashimshony et al. 2016).
DNA Damage Signaling Instructs Polyploid Macrophage Fate in Granulomas.
Specimen part, Cell line, Subject
View SamplesAn auxin-binding protein (Abp57) was previously isolated from rice and known to activate plasma membrane proton ATPase. The Abp57 function was characterised by overexpression in the rice and Arabidopsis. The transgene expression was driven by constitutive promoter, CaMV35S. Results from physiological experiments showed that the transgenic lines were tolerant to drought and salinity stress.
Microarray dataset of transgenic rice overexpressing <i>Abp57</i>.
Age, Specimen part
View SamplesWe used microarrays to detail the global program of gene expression during early hESC differentiation to mesendoderm using FBS, with and without RUNX1 depletion.
Transient RUNX1 Expression during Early Mesendodermal Differentiation of hESCs Promotes Epithelial to Mesenchymal Transition through TGFB2 Signaling.
Specimen part, Cell line
View SamplesWe identified 4,356 genes with expression differences associated with a high-fat diet, with 184 genes exhibiting a sex-by-diet interaction. Dietary fat dysregulated several pathways, such as cytokine-cytokine receptor interaction, chemokine signaling, and oxidative phosphorylation. Grant: Funding source: American Heart Association Grant number: 16PRE26420105 Title: The effect of maternal over-nutrition on obesity, epigenetics, and gene expression Awarded to Madeline Keleher Overall design: We performed RNA-seq in 21 total libraries, each with two mice of the same sex and diet pooled together (There were 6 low-fat-fed female libraries, 5 libraries of high-fat-fed females, 5 libraries of low-fat-fed males, and 5 libraries of high-fat-fed males). A 1x50 single read sequencing run was done on an Illumina HiSeq 2500 machine (Illumina Inc.)
A high-fat diet alters genome-wide DNA methylation and gene expression in SM/J mice.
Sex, Specimen part, Cell line, Subject
View SamplesStudy on selective vulnerability of certain brain regions to oxidative stress. Here we selected 4 brain regions (hippocampal CA1 and CA3, cerebral cortex, and cerebellar granular layer) to study this phenomenon.
Genomic and biochemical approaches in the discovery of mechanisms for selective neuronal vulnerability to oxidative stress.
Specimen part
View SamplesCoupling immunity and development is essential to ensure survival despite changing internal conditions in the organism. The metamorphosis of the fruit fly represents a striking example of drastic and systemic physiological changes that need to be integrated with the innate immune system. However, the mechanisms that coordinate development and immune cell activity in the transition from larva to adult in Drosophila remain to elucidate. The steroid hormone ecdysone is known to act as a key coordinator of metamorphosis. This hormone activates a nuclear receptor, the Ecdysone Receptor (EcR), which acts as a heterodimer with its partner Ultraspiracle (USP). Together, they activate the transcription of primary response genes, which in turn activate the transcription of a battery of late response genes. We have revealed that regulation of macrophage-like cells (hemocytes) by the steroid hormone ecdysone is essential for an effective innate immune response over metamorphosis. We have shown that in response to ecdysone signalling, hemocytes rapidly up regulate actin dynamics, motility and phagocytosis of apoptotic corpses, and acquire the ability to chemotax to damaged epithelia. Most importantly, individuals lacking ecdysone-activated hemocytes are defective in bacterial phagocytosis and are fatally susceptible to infection by bacteria ingested at larval stages, despite the normal systemic production of antimicrobial peptides. This decrease in survival is comparable to the one observed in pupae lacking immune cells altogether, indicating that ecdysone-regulation is essential to hemocyte immune functions and survival after infection.
Steroid hormone signaling is essential to regulate innate immune cells and fight bacterial infection in Drosophila.
Specimen part
View SamplesReprogramming intermediates (pre-iPSCs) were subjected to control DMSO, ascorbic acid (AA), 2i ( MAP kinase inhibitor + GSKinhibitor) or AA+2i conditions to assess conversion to the iinduced pluripotent stem cell state (iPSC) after 10days. Overall design: Pre-iPSC cells exposed to control DMSO, AA, 2i or AA+2i were harvested on day 2 for RNA sequencing after treatment, in biological triplicate.
Collaborative rewiring of the pluripotency network by chromatin and signalling modulating pathways.
No sample metadata fields
View SamplesGender dimorphism exists in the physiological response to diet and other environmental factors. Trans-hydrogenated fatty acid (TFA) intake is associated with an increase in coronary heart disease (CHD), and gender differences in the incidence of CHD are well documented. Neonatal administration of Monosodium Glutamate (MSG) causes stunted heart growth and hypoplasticity; and gender dimorphism at the growth hormone axis has been demonstrated in MSG-treated rodents. The identification of gender dimorphism in cardiac nutrigenomics may provide the basis for gender-specific medicine in the future.
Sex-dimorphism in cardiac nutrigenomics: effect of trans fat and/or monosodium glutamate consumption.
Sex, Specimen part
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