Project description:RNA-seq of IPEC-J2 cells samples from different groups to search for miRNA effect on gene expression. Lentivirus of miRNA 133, and miRNA 143 were used to knock down (KD), or overexpress (OV) the two miRNAs. LV3NC is the control for knock down, Lv20NC is control for overexpression. LV133 is the miRNA 133 knockdown, LV143 is the miRNA 133 knock down. OV143 is the miRNA 143 overexpression. miRNA 133 is no overexpression.
Project description:Deoxynivalenol (DON) frequently detected in a wide range of foods and feeds, inducing cytotoxicity to animals and humans. N6-methyladenosine (m6A) is an important epitranscriptomic marker with high abundance in eukaryotic mammals mRNAs. However, the role of the m6A methylomes in DON damage is still poorly understood. Here, we investigated the m6A transcriptome-wide profile in intestinal porcine epithelial cells (IPEC-J2) with and without 1000 ng/mL DON treatment via m6A sequencing and RNA sequencing. In total, 5406 new m6A peaks appeared with the disappearance of 2615 peaks in DON-induced IPEC-J2. The unique m6A-modified genes in DON-induced IPEC-J2 were associated with TNF signaling pathway. We identified 733 differentially expressed mRNA transcripts with hyper-methylated or hypo-methylated m6A peaks between DON-induced IPEC-J2 and normal IPEC-J2. Protein interaction network analysis and qPCR validation suggested that CSF2 probably acts as a promising new target for combating DON damage in IPEC-J2. Our first report of m6A transcriptome-wide map of IPEC-J2 cells presented here provides a starting roadmap for uncovering m6A functions that may affect DON infection.
Project description:To gain a more complete understanding of how porcine cathelicidin PR-39 influence the porcine intestinal epithelial cells, we profiled gene expression patterns in IPEC-J2 cell line in the presence or the absence of PR-39. two groups(control and PR-39),each group has three replicates
Project description:To understand the immunomodulatory effects of deoxyshikonin (a natural derivative of well-known Chinese medicine shikonin) on porcine intestinal epithelium cells, the transcriptome analysis was performed to explore the transcriptional profile of porcine IPEC-J2 cells after deoxyshikonin treatment.
Project description:The objective of this study is to investigate to what extent gene expression data of dietary interventions generated in IPEC-J2 in vitro model overlap with in vivo data. Gene expression was recorded in IPEC-J2 cells upon exposure to three different dietary interventions commonly used in livestock. In a further step, we compared the results with mucosal gene expression responses, as measured in animals exposed to the same compounds via the diet. As compounds we used zinc oxide, rye and the antibiotic amoxicillin. The GEO accession numbers of the in vivo datasets are provided in the paper "Enrichment of in vivo determined transcription data from dietary intervention studies with in vitro data provides improved insight into gene regulation mechanisms" (submitted to "Genes and Nutrition").
Project description:To gain a more complete understanding of how porcine cathelicidin PR-39 influence the porcine intestinal epithelial cells, we profiled gene expression patterns in IPEC-J2 cell line in the presence or the absence of PR-39.
Project description:The objective of this study was to investigate the effects of CDCA on the gene expression of IPEC-J2 cells. The control group (CON) treated with PBS and the CDCA group treated with both CDCA. Compared with the CON group, CDCA group could regulate the gene expression related proliferation genes.
Project description:Here we analysed different mechanisms of apical and basolateral deoxynivalenol (DON) toxicity reflected in the gene expression. We used the jejunum derived, polarized intestinal porcine epithelial cell line IPEC‑J2 as an in vitro cell culture model. Luminal and blood stream DON intoxication was mimicked by a DON application from the apical or basolateral compartment of ThinCert® membrane inserts for 72 h.
Project description:The molecular mechanisms involved in host pathogen interactions at mucosal surfaces needs to be better understood in order to develop immune-mediated methods of protection against pathogens. Cholera toxin (CT) has the rare ability for a protein of inducing robust mucosal immunity in the gut and is therefore an excellent model with which to determine mechanisms of adjuvanticity and immunogenicity at intestinal mucosal surfaces. Jejunal epithelial cells are one of the first sites of antigen encounter. Therefore a porcine intestinal epithelial cell line, IPEC-J2, was cultured in 6-well transwell plates in the presence or absence of 50 ng/ml cholera toxin for up to 8 hours and the cell layer was harvested for gene expression analysis using the Affymetrix porcine genome array and real-time PCR analysis. Affymetrix analysis identified, and real-time PCR analysis of 15 genes confirmed, an increase in gene expression for 59 genes and a decrease in gene expression for 14 genes under CT treatment. An 8 hour time course of expression revealed that by 2-4 hours after CT treatment, all 10 upregulated genes were differentially expressed and by 4-6 hours after CT treatment 3 of the 5 downregulated genes were differentially expressed. These data suggest that the potent mucosal adjuvanticity and immunogenicity of CT derives from rapid alterations in gene expression at the site of first antigen encounter with the immune system. Characterization of early immune gene expression may elucidate potential biological mechanisms for mucosal immune induction leading to the development of effective vaccines against enteric pathogens. Experiment Overall Design: Confluent IPEC-J2 cell monolayers were treated with or without 50 ng/ml cholera toxin in transwell dishes for 8h. The experiment was repeated 3 times for a total of 6 chips, 3 cholera toxin treated and 3 untreated. Gene expression was compared between cholera toxin treated and untreated cells.
Project description:The GeneChip Porcine Genome Array was used to identify the transcriptional response upon either Salmonella typhimurium (ST) or Salmonella choleraesuis (SC) infection in two porcine epithelial cell lines (IPEC-J2, from jejunum and IPI-2I, from ileum) during 2 and 4 hours post infection. The objectives in this study were first, to identify the different response between the epithelial cell lines from different gut regions; second, to study how the Salmonella serotypes used could elicit a different host response; and third, to determine the effect of the time-points on the differentially gene expression.