GSE162598
Next Generation Sequencing Facilitates Quantitative Analysis of Taurine Promoted Transcriptomes [Control+DON]
Purpose: DON inhibits the activation of gamadelta T cells. The goals of this study are to identify genes and pathways invovled in the regulation. Methods: Mouse splenic gamadelta T cells were stimulated with IL-23 and IL1ß, together with or without DON for 3 days. Then, the next-generation libraries of mRNA were prepared using VAHTS mRNA-seq v2 Library Prep Kit for Illumina® (Vazyme, Nanjing, China). The Library quality was determined by Bioanalyzer 4200 (Agilent, Santa Clara, CA, USA). Then the mRNA-seq libraries were sequenced in HiSeq ?10 system (Illumina, San Diego, CA, USA) on a 150bp paired-end run. The differentially expressed genes were selected as having more than 1 fold difference in their geometrical mean expression between the compared groups and a statistically significant p-value (<0.05) by analysis of DEseq2. The GO analysis on differentially expressed genes was performed with an R package: Clusterprofiler using a p<0.05 to define statistically enriched GO categories. Pathway analysis was used to determine the significant pathway of the differential genes according to Kyoto Encyclopedia of Genes and Genomes Database (http://www.genome.jp/kegg/), the enrichr database (https://maayanlab.cloud/Enrichr/) and DAVID Bioinformatics Resources 6.8 (https://david.ncifcrf.gov/). Results: Genes in the IL-23/STAT3 pathway were inhibited by taurine. As a result, the production of IL-17A decreased upon DON treatment. Overall design: MRNA profiles of IL-23+IL1ß stimulated and IL-23+IL-1ß plus DON treated gamadelta T cells were generated by deep sequencing, in triplicate, using illumina HiSeq ?10 system