Project description:Our aim was to identify Regenerating islet-derived (REG proteins in dogs. We annotated REG-family members in the canine genome and proteome, and identified one member of the REG3 subfamily, consequently termed REG3E by the Vertebrate Gene Nomenclature Committee. We verified REG3E gene expression using publicly available RNA-seq data (www.barkbase.org). We investigated the presence of REG3E protein in canine pancreatic tissue and plasma with Western blot and immunohistochemistry, using anti-human REG3A antibodies. Protein identity was subsequently confirmed with mass spectrometry on canine pancreas homogenates and plasma.
Project description:The expression of OCT2 was also increased in NK cells from Reg-1fl/fl in response to IL-12/18 stimulation implying that OCT2 has an intrinsic function not only in Regnase-1-deleted NK cells but also in wild-type NK cells. To identify the genome-wide binding sites of OCT2, we performed chromatin immunoprecipitation sequencing (ChIP-seq) for OCT2 in Reg-1fl/fl- and Reg-1ΔNK-NK cells.
Project description:Analysis of the maize alternative splicing landscape, including transcript discovery and mapping of genotype-dependent variations in alternative splicing using B73, Mo17 and the SX19 inbred mapping population Total RNA was isolated from 5 week old leaves of hydroponically grown maize plants and used to construct RNA seq libraries
Project description:The innate immune system can recognize tumor cells and coordinate with the adaptive immune system to initiate an anti-tumor response within the tumor microenvironment. To investigate the role of NK cells and their interaction with other immune cells in the anti-tumor immunity in Reg-1ΔNK. We subcutaneously injected B16F10 cells into Reg-1fl/fl and Reg-1ΔNK, followed by scRNA-seq analysis of the whole tumor cell suspension and sorted TIL-NK cells.
Project description:By using oligonucleotide microarray-based technology as a tool for rapid mapping of unique differences in gene expression, we found that inhibiting serpin B13 in BALB/c mice treated with a mAb according to our protocol resulted in the upregulation of relatively few genes, including several Reg genes, which have been implicated in β-cell proliferation.
Project description:The purpose of this data set is to determine if bradyzoites impact the host transcripton in a similar manner as tachyzoites. We found that 3853 differentially expressed genes (DEGs) are shared between tachyzoite infected cells and bradyzoite infected cells (41%; 2161 up-reg., 1692 down-reg.) when compared to uninfected host samples, leaving 3356 tachyzoite specific DEGs (1488 up-reg., 1868 down-reg.), and 1951 DEGs specific to bradyzoite infection (1075 up-reg., 876 down-reg). We conclude therefore that bradyzoites manipulate the host cell and that this is unique when compared to tachyzoites.
Project description:Differential expression was used to access gene differences after Entamoeba histolytica infection. Entamoeba histolytica is an important diarrheal pathogen worldwide, and induces apoptosis of the intestinal epithelium as part of its disease process. Regenerating (REG) 1 protein is anti-apoptotic. We investigated the involvement of REG 1 in E. histolytica colitis. Colonic biopsy samples were obtained from 8 subjects with acute E. histolytica colitis, and again 60 day later during convalescence. Gene expression in the human colon during acute and convalescent E. histolytica disease was evaluated using microarray and confirmed by polymerase chain reaction (PCR). REG 1 protein expression was evaluated with immunohistochemistry. The mechanism of REG 1 involvement in E. histolytica disease was subsequently investigated with a mouse model. REG 1A and REG 1B were the most upregulated genes in the human intestine in acute versus convalescent E. histolytica disease (p=0.003 and p=0.006 respectively). PCR confirmed the microarray results (p=<0.001 and p=0.001 respectively). Increased REG 1A and REG 1B protein expression was similarly observed by immunohistochemistry. REG 1 -/-mice were found to be significantly more susceptible to E. histolytica infection than wild type mice.
Project description:Having identified the crucial role of IFN-γ in anti-tumor immunity of Reg-1ΔNK, we attempted to elucidate the mechanism underlying the regulation of Ifng in the context of Regnase-1 deletion. Despite the notable increase in Ifng mRNA expression in Reg-1ΔNK-NK cells, Ifng was not a direct substrate of Regnase-1. However, the production of the IFN-γ was further enhanced upon stimulation with IL-12/18. We proposed that modifications may have occurred in the transcriptional regulation of Ifng in Reg-1ΔNK-NK cells.
Project description:Variability of regenerative potential among animals has long perplexed biologists. Based on their amazing regenerative abilities, planarians have become important models for understanding the molecular basis of regeneration; however, planarian species with limited regenerative abilities are also found. Despite the importance of understanding the differences between closely related, regenerating and non-regenerating organisms, few studies have focused on the evolutionary loss of regeneration, and the molecular mechanisms leading to such regenerative loss remain obscure. Here we examine Procotyla fluviatilis, a planarian with restricted ability to replace missing tissues, utilizing next-generation sequencing to define the gene expression programs active in regeneration-permissive and regeneration-deficient tissues. We found that Wnt signaling is aberrantly activated in regeneration-deficient tissues. Remarkably, down-regulation of canonical Wnt signaling in regeneration-deficient regions restores regenerative abilities: blastemas form and new heads regenerate in tissues that normally never regenerate. This work reveals that manipulating a single signaling pathway can reverse the evolutionary loss of regenerative potential. RNA-seq experiments to identify gene expression changes following amputation in body regions with variable regenerative potential. Adult Procotyla fluviatilis were amputated at sites either anterior or posterior to the pharynx. After 24 hours post-amputation, tissues near the amputation site were excised and RNA was extracted. Similar tissues were excised from uncut control animals. Samples were processed for RNA-seq using Illumina procedures. We generated a de novo P. fluviatilis transcriptome and used RNA sequencing (RNA-seq) to characterize transcripts from excised tissue fragments in Reg+ and Reg- body regions 24 hours post-amputation. We performed parallel analyses on tissues excised from intact animals at identical body regions to account for regional differences in transcripts, thereby identifying changes resulting from amputation. Samples A1-A3 = Regeneration-proficient (Reg+) tissue excision 24 hours after amputation. Samples B1-B3 = Tissue excision from regeneration-proficient (Reg+) region but not amputated. Samples C1-C3 = Tissue excision from regeneration-deficient (Reg-) tissues 24 hours after amputation. Samples D1, D3-D4 = Tissue excision from regeneration-deficient (Reg-) region that was not amputated.