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Epigenetic silencing of antiviral genes renders clones of Huh-7 cells permissive for hepatitis C virus replication.
Specimen part
View SamplesMX1 is a well-characterized interferon-induced antiviral gene. MX1 is activated by viral infection due to interferon production in cells. We treated non-permissive Huh7 cells and permissive HRP4 cells with interferon. We compared the expression of genes induced by interferon to determine host factors affecting HCV replication.
Epigenetic silencing of antiviral genes renders clones of Huh-7 cells permissive for hepatitis C virus replication.
Specimen part
View SamplesWe FACS-isolated single thirst-associated neurons from the median preoptic hypothalamus of mice and determined their individual transcriptomes. This characterization revealed a molecularly distinct population of excitatory thirst-associated neurons that is responsible for producing thirst motivational dirve. Overall design: Thirst-associated cells in the preoptic hypothalamus of mice were labeled using the Fos-p2A-CreER; Ai14 reporter mouse after 48 hour water deprivation. The preoptic hypothalamus was dissociated, and individual tdTomato+ cells were sorted into 96 well plates with lysis buffer. The transcriptomes of 570 putative cells were amplified using SmartSeq2 single-cell RNA-seq (Picelli et al., 2014). Libraries were prepared using an Illumina Nextera XT following Illumina''s protocols, and sequenced on an Illumina NextSeq 500. Data were processed using RSEM, and mapping directly to the ENSEMBL transcriptome, and quantified at a per-transcript level in TPM units. FASTQ files containing few reads (<1 MB in size for first paired-end read) were not mapped or subsequently analyzed; final dataset comprises 505 cells.
Thirst-associated preoptic neurons encode an aversive motivational drive.
Specimen part, Cell line, Subject
View SamplesDrugs directly targeting Hepatitis C (HCV) are often rendered useless by the high mutation rate of the virus. Thus, we deduce that targeting of host factor that affect HCV replication may provide enhanced therapy fort HCV infection. Hepatocyte cell line Huh7 is known to be non-permissive for Hepatits C (HCV) replication. Through a method developed by the Rice laboratory (Blight, K.J., et al., J Virol, 2002), selection of a small subset of permissive hepatocytes is possible. The Rice laboratory generated the first permissive cell line, Huh7.5, using this method. We generated another permissive cell line, HRP1, using the same method.
The membrane-bound transcription factor CREB3L1 is activated in response to virus infection to inhibit proliferation of virus-infected cells.
Specimen part, Cell line
View SamplesMembrane-bound transcription factor CREB3L1 undergoes Regulated Intramembrane Proteolysis (RIP) in response to Hepatitis C infection. RIP activates CREB3L1 so that it can prevent the growth of HCV infected cells through the action of downstream genes. We over-expressed a truncated form of CREB3L1 that does not require RIP to enter the nucleus. Cells over-expressing this truncated form were isolated by Fluorescence Activated Cell Sorting (FACS).
The membrane-bound transcription factor CREB3L1 is activated in response to virus infection to inhibit proliferation of virus-infected cells.
No sample metadata fields
View SamplesMouse skin fibroblasts (MSFs) were obtained from a FASST (Fibroblasts Accelerate Stromal-Supported Tumorigenesis) mouse. This mouse model allows for spatial and temporal control for senescence induction by using a stromal specific Cre-recombinase driven by the pro-collagen-alpha II promoter. The stromal specific Cre activates expression of the p27IRESGFP transgene that is expressed from the ROSA locus. We cultured the MSFs in vitro, induced senescence using 10uM tamoxifen added to the media. Non-senescent cells were treated with equal volume of vehicle alone (ethanol). Upon tamoxifen treatment, cells were moved to a modular incubation chamber and maintained at 3% oxygen at 37 degrees celcius for 12 days total before collection. At the time of collection, cells were trypsynized and pelleted by centrifugation. The cells were lysed using Trysol reagent and RNA was isolated using a RiboPure RNA isolation kit (Ambion). Overall design: For this study, 2 treatment groups were analyzed (non-senescent, EtOH samples and senescent, TAM samples). Each treatment group was performed 3 times for a total of 6 samples for analysis. The gene expression analysis is a comparison of expression in senescent (TAM) vs non-senescent (EtOH) mouse skin fibroblasts.
Stromal senescence establishes an immunosuppressive microenvironment that drives tumorigenesis.
Specimen part, Cell line, Treatment, Subject
View SamplesOptic nerves are an accessible part of the CNS, providing a source of glia without the presence of neuronal cell bodies. Therefore, an analysis was carried out of gene expression in optic nerves at P4, before myelination begins and at P10, when myelination is very actively proceeding. The goal was to obtain a profile of the changing gene expression that accompanies this transition from unmyelinated CNS nerve to myelinated nerve.
Towards resolving the transcription factor network controlling myelin gene expression.
Specimen part
View SamplesTo better understand the impact of integrin beta3 signaling in myeloid cells on the tumor microenvironment, we compared the gene expression profiles of FACS isolated GFP+ PyMT-BO1 MFP tumor cells and also M2 TAMs (CD11b+Gr1-F4/80+CD206+) from tumor tissue of WT mice and b3 mice.
Antagonizing Integrin β3 Increases Immunosuppression in Cancer.
Specimen part
View SamplesThe intestinal immune system must elicit robust immunity against harmful pathogens but restrain immune responses directed against commensal microbes and dietary antigens. The mechanisms that maintain this dichotomy are poorly understood. Here we describe a population of CD11b+F4/80+CD11c macrophages in the lamina propria (LP) that express several anti-inflammatory molecules including interleukin 10 (IL-10), but little or no pro-inflammatory cytokines, even upon stimulation with Toll-like receptor (TLR) ligands. These macrophages induced, in a manner dependent on IL-10, retinoic acid and exogenous transforming growth factor-, differentiation of FoxP3+ regulatory T cells. In contrast, LP CD11b+ dendritic cells elicited IL-17 production. This IL-17 production was suppressed by LP macrophages, indicating that a dynamic interplay between these subsets may influence the balance between immune activation and tolerance.
Lamina propria macrophages and dendritic cells differentially induce regulatory and interleukin 17-producing T cell responses.
No sample metadata fields
View SamplesActivation of inflammatory pathways in human IBD
Activation of an IL-6:STAT3-dependent transcriptome in pediatric-onset inflammatory bowel disease.
No sample metadata fields
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