In the current study, we investigated the collective roles of protein tyrosine phosphatases (PTPs) and histone deacetylases (HDACs) on regulation of IRG expression in human choriocarcinoma cells by genome-wide transcriptional profiling. Logic-rules were optimized to derive rules governing gene expression patterns observed upon different combinations of treatment with PTP and HDAC inhibitors. The data reveal that IRGs can be divided into distinct subsets that are differentially modulated by co-treatment of Jar cells with IFN-? and PTP versus HDAC inhibitors, respectively. Furthermore, promoter analysis of the genes governed by the rules identifies transcription factor binding sites associated with the different gene subsets. Thus, the regulatory modes identified in this study provide insights into the complex regulation of inflammatory pathways at the fetal-maternal interface, as well as mechanisms that choriocarcinoma cells may utilize to promote their survival.
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View SamplesRetinoic acid-inducible gene-I (RIG-I) is a cytosolic pathogen recognition receptor that initiates the innate immune response against many RNA viruses. RIG-I also has been shown to sense some DNA viruses, and host RNA polymerase III (RNA Pol III), a cytosolic DNA sensor, converts cytosolic AT-rich DNA into RNA to be sensed by RIG-I. We previously showed that the RIG-I restricts Kaposi Sarcoma-associated herpesvirus (KSHV) reactivation (J Virol. 2014 May;88(10):5778-87). In this study, we report that KSHV stimulates the RIG-I signaling pathway in an RNA Pol III-independent manner and subsequently induces type I IFN responses. Knockdown or inhibition of RNA Pol-III had no effect on IFN-ß induction by KSHV. By using CLIP (Cross-Linking and Immunoprecipitation) and RNA deep sequencing technologies, we identified multiple KSHV regions that give rise to RNA fragments binding to RIG-I, such as ORF810420-10496, ORF6411058-110675, Repeat region (LIR1)119059-119204, and ORF2543561-43650. The sequence dissimilarity between these fragments suggests that RIG-I detects a particular structure rather than a specific sequence motif. Synthesized ORF810420-10496 RNA stimulated RIG-I-dependent but RNA Pol III-independent IFN-ß signaling. In summary, some KSHV viral RNAs are sensed by RIG-I in an RNA Pol III-independent manner.
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View SamplesFresh splenic Treg cells (CD4+CD25+YFP+) were isolated from 6-week-old Senp3+/+Foxp3-Cre and Senp3fl/flFoxp3-Cre mice and stimulated with anti-CD3 and anti-CD28 for 24 hours. Activated Treg cells were used for total RNA isolation with TRIzol and subjected to RNA-sequencing.
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View SamplesSingle NG2-glia with GFP fluorescence labeling from PDGFRaCreER; mGFP mice was selected and aspirated into a glass pipette from hippocampal acute slices. In brief, cells were picked promptly by micromanipulation and immediately placed in lysis buffer. All NG2 glial cells were collected within 3 h after slice preparation. The selected NG2-glia were processed for single-cell RNA extraction and reverse transcription within 1 h and were subjected to RNA-sequencing.
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Sex, Specimen part, Cell line
View SamplesB cell mRNA profiles of 18-week-old wild type (WT) and B cell-specific SHIP1 knockout (SHIP?B) mice were generated by deep sequencing, in duplicate, using Illumina Nextseq500.
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View SamplesRNA-Seq analysis was used to study the profile of CD8 t cells from melanoma patients before and after treatment to detect transcriptional changes in peripheral blood
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View SamplesActively proliferating cells must constantly monitor and re-adjust their metabolic pathways to ensure phospholipid homeostasis for the replenishment of membranes and intracellular trafficking. Multiple studies have suggested that the lysine acetyltransferase NuA4 has a role in fine-tuning phospholipid metabolism in Saccharomyces cerevisiae, however the role of NuA4 in phospholipid homeostasis remains poorly defined. NuA4 mutants have increased gene expression of inositol-3-phosphate synthase, INO1 and overproduce inositol. NuA4 mutants are also display synthetic sickness with a mutant of the lipid remodeling gene SEC14. Here using a combination of genetics and transcriptional profiling, we explore the connections between NuA4, inositol and Sec14. We found that NuA4 mutants exacerbated the inositol auxotrophy of sec14-1ts. Transcriptome studies reveal that loss of the NuA4 subunit EAF1 in sec14-1ts depresses INO1 expression but not all inositol/choline responsive phospholipid genes. This suggests eaf1? cells are defective in coordinating phospholipid homeostasis beyond inositol production. In fact, we discovered that eaf1? cells have significantly lower lipid droplet levels and that inhibition of the fatty acid biosynthesis pathway increased the growth defect of sec14-1ts to a similar extent as untreated sec14-1tseaf1?. Altogether, our work identifies a role for NuA4 as a critical mediator of phospholipid metabolism, potentially as positive regulator of fatty acid biosynthesis.
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Specimen part, Disease, Cell line
View SamplesComparison with antibiotic susceptible and multi-drug resistant Pseudomonas aeruginosa, and responses to antibiotic stresses in multi-drug resistant Pseudomonas aeruginosa
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Specimen part, Cell line
View SamplesMalaria, caused by Plasmodium parasites is responsible for the illness of millions of individuals each year. Plasmodium sporozoites inoculated by mosquitoes migrate to the liver and infect hepatocytes prior to release of merozoites that initiate symptomatic blood-stage malaria. Parasites are thought to be restricted to hepatocytes throughout this obligate liver-stage of replication and differentiation. In contrast to this notion, we found that a subset of hepatic CD11c+ cells co-expressing F4/80, CD103, CD207 and CSF1R, acquired a substantial parasite burden during the liver-stage of malaria, but only after initial hepatocyte infection. These CD11c+ cells found in the infected liver and liver-draining lymph nodes exhibited transcriptionally and phenotypically enhanced antigen-presentation functions; and primed protective CD8 T cell responses against Plasmodium liver-stage restricted antigens. Our findings uncover a novel aspect of Plasmodium biology as well as the fundamental mechanism by which CD8 T cell responses are primed against liver-stage malaria.
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