RNA-Seq of the aorta of hypercholesterolemic mice treated by either ticagrelor, clopidogrel, or control.
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ABSTRACT: We fed hypercholesterolemic mice high fat diet containing either ticagrelor, clopidogrel, or nothing (control) for 16 weeks, isolated the aortae, extracted RNA, and subjected the RNA to RNA-Seq using NGS.
Project description:We fed hypercholesterolemic mice high fat diet containing either ticagrelor, clopidogrel, or nothing (control) for 16 weeks, isolated the liver, extracted RNA, and subjected the RNA to RNA-Seq using NGS.
Project description:We sequenced the transcriptome of 34 single olfactory sensory neurons (OSNs) manually picked from OMP-GFP mice, ensuring the cells expressed high levels of GFP, a marker of mature OSNs. We pooled the olfactory mucosa from 2 male and 2 female animals, to obtain single-cell suspensions for cell picking. We only picked healthy-looking cells. From each, we constructed libraries for single-cell RNA-sequencing, to characterise the diversity of OR genes expressed.
Project description:To determine the role of Ascl1 in beta cell development, function, and metabolic stress response, we generated beta cell specific Ascl1 knockout mice and assessed their glucose homeostasis, islet morphology, and gene expression after feeding a normal diet, a high fat diet (HFD) for 12 weeks, or on a background of Abcc8 (KATP channel subunit) knockout mice. For the RNA-seq analysis, islets from male Ascl1betaKO and littermate control mice (N = 4 for each genotype and condition) were collected from three different conditions: 1) normal diet fed 2) HFD fed for 12 weeks, 3) on a background of homozygous Abcc8 allele.
Project description:New mechanisms-of-action of anthocyanins (ACNs) provided by a red-fleshed apple compared with a white-fleshed apple ACN-poor, and with an ACN-rich extract on the proteome profile of aorta and heart as cardiovascular key tissues were determined. Hypercholesterolemic Wistar rats were separated into the corresponding groups to analyze the proteomic profile of the aorta and heart tissues using nano-liquid chromatography coupled to mass-spectrometry. Red-fleshed apple downregulated CRP, C1QB and CFP related-inflammation. White-fleshed apple reduced C1QB, CFB, CFD, C3, and C9 related to the complement system, reduced MB and CP related to iron metabolism, and increased ME1, PKM, and PC related to energy homeostasis. ACN-rich extract increased FMOD, TAGLN, and CAP1 related to cellular structure and decreased PRKACA, IQGAP1, and HSP90AB1 related to cellular signaling. Red-fleshed apple rich in ACNs suggested an anti-inflammatory effect while white-fleshed apple reduced the complement system protein-related. An apple matrix effect reduced inflammatory proteins regardless their ACN content.
Project description:This study aimed to identify small molecules that inhibit vascular calcification with computational approach. To examine the mechanism of calcification inhibition, proteomics analysis was performed using aorta tissue samples to clarify therapeutic target.
Project description:CTCF is a highly conserved and ubiquitously expressed protein involved in several fundamental processes such as fine-tuning gene expression, imprinting, X-chromosome inactivation and 3D chromatin organisation. To understand the impact of differences in the concentration of CTCF abundance on these processes, we exploit a CTCF hemizygous mouse model with a stable reduction in the concentration of this protein. We derived twelve independent primary lines of mouse embryonic fibroblasts (MEFs) from six wildtype and six CTCF-hemizygous mouse E13.5 embryos. Total RNA from each MEF line was purified using QIAzol Lysis Reagent (Qiagen); DNase treatment and removal was performed using the TURBO DNA-freeTM Kit (Ambion, Life Technologies). Libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina) and sequenced in an Illumna HiSeq4000 to produce 150bp paired-end reads. On the same MEF lines we have performed ChIPseq for CTCF, H3K4me3 and H3K27ac and HiC.
Project description:Diet-induced obesity is reported to induce a phenotypic switch in adipose tissue macrophages from an antiinflammatory M2 state to a proinflammatory M1 state. Telmisartan, an angiotensin II type 1 receptor antagonist and a peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist, reportedly has beneficial effects on insulin sensitivity. We studied the effects of telmisartan on the adipose tissue macrophage phenotype in high fat-fed mice. Telmisartan was administered for 5 weeks to high fat-fed C57BL/6 mice. Insulin sensitivity, macrophage infiltration, and the gene expressions of M1 and M2 markers in epididymal fat tissues were examined. Insulin- or a glucose-tolerance test showed that telmisartan treatment improved insulin resistance, decreasing the body weight gain, visceral fat weight and adipocyte size without affecting the amount of food intake. Telmisartan treatment reduced the number of CD11c-positive cells and crown-like structures. Telmisartan reduced the mRNA expressions of M1 macrophage markers, such as TNF-alpha and IL-6, and increased the expression of M2 markers, such as IL-10 and Mgl2. The reduction of M1 macrophage markers, as well as the increased gene expression of M2 markers especially IL-10, is a possible mechanism for the improvement of insulin sensitivity by telmisartan. Six-week-old male C57BL/6J mice were purchased from CLEA Japan. The mice were fed a chow that contained 10% of its calories from fat (control) or a high-fat diet (HFD) that contained 30% of its calories from fat for 24 weeks. The high fat-fed mice were randomized to 3 groups. Either telmisartan (~3 mg/kg/day) in drinking water (HFD+Tel), candesartan (~3 mg/kg/day) in drinking water (HFD+Can), or a HFD without any drugs (HFD) was administered for the next 5 weeks. Two mice were treated per group. Epididymal adipose tissues were rapidly removed from each mouse. Gene expression in epididymal fat tissue was analyzed using a GeneChip® system with the Mouse Genome 430 2.0 Array, which was spotted with 45,101 probe sets (Affymetrix, Santa Clara, CA, USA). Sample preparation for the array hybridization was performed according to the manufacturer’s instructions. In short, 5 μg of total RNA was used to synthesize double-stranded cDNA using the GeneChip® Expression 3′-Amplification Reagents One-Cycle cDNA Synthesis Kit (Affymetrix). Biotin-labeled cRNA was then synthesized from the cDNA using GeneChip® Expression 3′-Amplification Reagents for IVT Labeling (Affymetrix). After fragmentation, the biotinylated cRNA was hybridized to arrays at 45 °C for 16 h. The arrays were washed, stained with streptavidin-phycoerythrin, and scanned using a probe array scanner. The scanned chip was analyzed using the GeneChip Analysis Suite software (Affymetrix). Hybridization intensity data were converted into a presence/absence call for each gene, and changes in gene expression between experiments were detected by a comparison analysis. Data was shown as the fold change relative to the expression level of normal chow-fed mice.
Project description:Glioblastoma (GBM) remains among the deadliest of human malignancies, and the emergence of the cancer stem cell (CSC) phenotype represents a major challenge to durable treatment response. Because the environmental and lifestyle factors that impact CSC populations are not clear, we sought to understand the consequences of diet on CSC enrichment. We evaluated disease progression in mice fed an obesity-inducing high-fat diet (HFD) versus a low-fat, control diet. HFD resulted in hyper-aggressive disease accompanied by CSC enrichment and shortened survival. HFD drove intracerebral accumulation of saturated fats, which inhibited the production of the cysteine metabolite and gasotransmitter, hydrogen sulfide (H2S). H2S functions principally through protein S-sulfhydration and regulates multiple programs including bioenergetics and metabolism. Inhibition of H2S increased proliferation and chemotherapy resistance, whereas treatment with H2S donors led to death of cultured GBM cells and stasis of GBM tumors in vivo. GBM specimens present an overall reduction in protein S-sulfhydration, primarily associated with proteins regulating cellular metabolism. These findings provide new evidence that diet modifiable H2S signaling serves to suppress GBM by restricting metabolic fitness, while its loss triggers CSC enrichment and disease acceleration. Interventions augmenting H2S bioavailability concurrent with GBM standard of care may improve outcomes for GBM patients.
Project description:Both multiple myeloma (MM) and systemic lupus erythematosus (SLE) are characterized with abnormal production of plasma cells. In both diseases, the process of B cells differentiate into plasmablast/plasma cell is disordered. Despite the continuous research on the development of prognostic factors and introduction of new agents, dysregulation of plasmablast/plasma cells in MM and SLE is still uncontrolled. Thus, it is necessary to explore the novel therapeutic target to plasmablast/plasma cells. Because of plasmablast-like, Mus musculus myeloma SP 2/0 cell line was selected to explore the novel therapeutic target to plasmablast/plasma cells. To explore the novel therapeutic target to plasmablast/plasma cells, we determined mRNA profiles in SP 2/0 cells compared to naive B cells by RNA-seq. RNA-seq was done with an Illumina HiSeq 2500 instrument at GENEWIZ, Suzhou, China.
Project description:The aim of the experiment was to analyze key molecular changes using an unbiased, transcript-based approach in an experimental obesity model. Small expression RNA sequencing analysis was used to reveal expression changes. Male Long-Evans rats were given a control (CON) or high fat (20%) high sucrose (15%) diet (HFS) for 25 weeks. From week 16, the animals were injected. 0.25 mg · kg - 1 daily with selegiline (CON + S and HFS + S) or vehicle (CON, HFS). Left ventricular samples from all four groups (n = 4-5) were analyzed.