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Application of Chromatrap® Solid-State Chromatin Immunoprecipitation (ChIP) for epigenetic profiling of insects

ABSTRACT: Application of Chromatrap® Solid-State Chromatin Immunoprecipitation (ChIP) for epigenetic profiling of insects

PROVIDER: PRJNA498662 | ENA |

REPOSITORIES: ENA

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Application of Chromatrap® Solid-State Chromatin Immunoprecipitation (ChIP) for epigenetic profiling of insects

Project description:We report the application of Chromatraps® Solid-State Chromatin Immunoprecipitation technology for epigenetic profiling of histone modifications in insects. Here, we present the optimised protocol and conditions of Chromatrap® kits for successful ChIP and high-throughput sequencing of established model species, Drosophila melanogaster and the emerging model for behavioural plasticity Nicrophorus vespilloides. We highlight successful ChIP-seq of Drosophila melanogaster (Oregon-R) of comparable quality to modENCODE data and present successful enrichment of histone marks for Nicrophorus vespilloides. The addition of this insect-based ChIP-seq protocol provides a set of optimal guidelines to aid streamline end-users epigenetic research focus and reduce experimental time.

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IL-36 signaling in keratinocytes controls early IL-23 production in Aldara®-induced skin inflammation in mice

Project description:IL-36R signaling plays an important role in the pathogenesis of psoriasis. We ought to assess the specific function of IL-36R in keratinocytes for the pathology of Aldara®-induced psoriasis-like skin inflammation. Il36r-/- (KO) and Il36rΔK (DK) mice as well as their littermate controls, respectively Il36r+/+ (WT) and Il36rfl/fl (FL) mice, were challenged with the topical application of Aldara® on one ear during 3 or 7 days. RNASeq was performed on treated (AL) and untreated (CT) ears. Fold change analysis combined with RPKM and RT-qPCR allowed us to identify genes which were specifically regulated by IL-36R signaling in keratinocytes during the course of the treatment. We notably demonstrated that both IL-23 subunits (Il23a and Il12b) fell into this category.

2020-04-23 | GSE143688 | GEO

Application of the PM6 method to modeling the solid state.

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IL-7-primed bystander CD8 tumor-infiltrating lymphocytes optimize the antitumor efficacy of T-cell engager immunotherapy in solid tumors

Project description:Bispecific T-cell engagers (TCEs) show promising clinical efficacy in blood tumors, but their application to solid tumors with immunosuppressive environments remains challenging. Here, we show that Fc-fused IL-7 (rhIL-7-hyFc) changes the intratumoral CD8 T cell landscape, allowing TCE-immunotherapy to function effectively in solid tumors. In order to explore the transcriptomic changes induced by rhIL-7-hyFc or a combination of rhIL-7-hyFc and TCE treatment at the clonal level, we performed scRNA/TCR-seq.

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Confirmation of SOMAmer® Enrichment from Biological Matrices using Mass Spectrometry

Project description:In an effort to confirm binding of SOMAmer®s to their respective targets in an endogenous matrix, we have conducted a series of experiments using SOMAmer®s to enrich target proteins followed by measurement using two mass spectrometry techniques: data dependent analysis (DDA) and multiple reaction monitoring (MRM). For data dependent analysis, the library of 4783 SOMAmer®s were multiplexed in sets of 8 and used for enrichment of target proteins from cell lysate and conditioned media, as well as human plasma and serum. A subset of the SOMAmer®s were screened in urine by this global profiling technique as well. Cell lines were selected from Cancer Cell Line Encyclopedia for screening by comparing gene expression of the target proteins measured by RNA sequencing across the CCLE.1 Using the criterion of FPKM value greater than 5 for the target protein, the minimum number of cell lines was grown to maximize protein coverage across the SOMAmer® library. For a given cell line, the presence of at least 8 target proteins with FPKM values greater than 5was required for inclusion. Approximately 400 target proteins were not covered by this strategy were screened in serum and plasma only. For enrichment from biological matrices, SOMAmer®s were combined into sets of 8 such that the potential interaction with similar proteins or binding partners was minimized. Non-specific binding of the SOMAmer®s was assessed in each matrix by using a SOMAmer® generated against the bacterial protein CysH. Target protein spectral counts in the SOMAmer® enriched samples were compared to the respective CysH control. A positive hit is defined as target protein detection with a minimum of 2 spectral counts and signal over CysH background greater than 10X (if applicable).

2018-08-03 | PXD008819 | Pride

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