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The transcriptome data of immature dendritic cells were treated with hydrogen peroxide

ABSTRACT: RNA sequencing(seq-RNA) was used to analyze the underlying mechanism of immature dendritic cells (imDCs) response to extracellular reactive oxygen species (ROS). The oxidative stress model of imDCswas constructed by simulating ROS with H2O2. The final H2O2 concentration was determined by CCK8 and Annexin V-FITC/ PI double staining methods. The free migration of imDCs was analyzed by cell real-time imaging. Total RNA was extracted and transcriptome sequencing was performed. Go and KEGG enrichment analysis of differential expressed genes(DEGs) was performed. Transcripts related to metabolism and skeleton regulation were screened according to the changes of differential gene expression ≥1 and FDR≤0.05. The result showed that there were no significant changes in cell viability and apoptosis of imDCs when H2O2 concentration was less than 100 μmol/L. The free migrate ability of imDCs was enhanced in ROS environment. RNA-seq data showed that 282 differentially expressed genes between the control group and the H2O2 treatment group. Among them, 225 differentially expressed genes were down-regulated, and 57 were up-regulated. GO and KEGG enrichment analysis found that these genes were involved in metabolism, cell cycle and cytoskeleton related signaling pathway. Gsta1, Gsta3，Cstdc5，Cstdc5 and Prdx1 involved in response to xenobtiotic metabolism were up-regulated. S100a8, Tm4sf19 and Ppbp involved in skeleton regulation were also up-regulated. While Nusap1, Stmn1, Kif20 and Prc1 were down-regulated. This study suggested that the activation of antioxidant system, cytoskeletal remodeling and cell cycle arrest may be involved in regulating the migration ability of imDCs in response to extracellular ROS.

ORGANISM(S): Mus musculus

PROVIDER: GSE269439 | GEO | 2024/06/16

REPOSITORIES: GEO

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Project description:Aliivibrio salmonicida causes âcold-water vibriosisâ (or âHitra diseaseâ) in fish, including marine-reared Atlantic salmon. During development of the disease the bacterium will encounter macrophages with antibacterial activities such as production of damaging reactive oxygen species (ROS). To defend itself the bacterium will presumably start producing detoxifying enzymes, reducing agents, and proteins involved in DNA and protein repair systems. Even though responses to oxidative stress are well studied for a few model bacteria, little work has been done in general to explain how important groups of pathogens, like members of the Vibrionaceae family, can survive at high levels of ROS. We have used bioinformatic tools and an âomics approach to study how A. salmonicida responds to hydrogen peroxide (H2O2). First, we used the recently published genome sequence to predict potential binding sites for OxyR (H2O2 response regulator). The computer-based search identified OxyR sites associated with 20 single genes and 8 operons, and these predictions were compared to experimental data from Northern blot analysis, microarray analysis and 2D gel electrophoresis. In general, OxyR binding site predictions and experimental results are in agreement. Up- and down-regulated genes are distributed among all functional gene categories, but a striking number of â¥2 fold up-regulated genes encode proteins involved in detoxification or DNA protection and repair, are part of reduction systems, or are involved in carbon metabolism and regeneration of NADH/NADPH. Our predictions and âomics data corroborates well with findings from other model bacteria, but also suggest species-specific gene regulation. Two-condition experiment, cells grown in LB medium (control samples) vs. cells grown under oxidative stress (H2O2) (stimulated samples). Samples collected from three different timepoints (15, 30 and 60 min). Technical replicates for each timepoint: 3 control, 3 stimulated, independently grown and harvested. One replicate per array.

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The transcriptome data of immature dendritic cells were treated with hydrogen peroxide

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