Project description:Purpose: The goals of this study are to compare NGS-derived 3T3-L1-NC transcriptome profiling (RNA-seq) to LIGHT overexpression 3T3-L1 cells and find out the DEGs Methods: mRNA profiles of 3T3-L1-NC and 3T3-L1-LIGHT before and after differentiation into beige adipocytes were generated by deep sequencing, in triplicate, using Illumina HiSeq. The sequence reads that passed quality filters were analyzed at the transcript isoform level. Results: Using an optimized data analysis workflow, we mapped about 6.53 Gb data per sample. The average genome mapping rate is 87.27% and the average gene mapping rate is 80.18%. 18,255 genes were identified in which 17,367 of them are known genes and 1,001 of them are novel genes. 13,086 novel transcipts were identified in which 9,304 of them are previously unknown splicing event for known genes, 1,001 of them are novel coding transcripts without any known features, and the remaining 2,781 are long noncoding RNA. Conclusions: Our study represents the first detailed analysis of the impact of LIGHT on gene expression in process of beige adipocytes biogenesis with biologic replicates, generated by RNA-seq technology.
Project description:Next Generation Sequencing Facilitates Quantitative Analysis of the control 3T3-L1 (3T3-L1-NC) and LIGHT (tnfsf14) overexpression 3T3-L1 cells (3T3-L1-LIGHT) before and after differentiation into beige adipocytes
Project description:We analyzed RING1B binding regions in 3T3-L1 cell lines transduced with the retroviral vector for V5-tagged Fbxl10 or its dF-box mutant. RING1B ChIP-seq in empty, Fbxl10-1, and dF-box mutant vector transduced 3T3-L1 preadipocytes, in duplicate, and V5-Fbxl10 ChIP-seq in Fbxl10 overexpressing 3T3-L1 cells
Project description:Histones were isolated from brown adipose tissue and liver from mice housed at 28, 22, or 8 C. Quantitative top- or middle-down approaches were used to quantitate histone H4 and H3.2 proteoforms. See published article for complimentary RNA-seq and RRBS datasets.