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Learn MorePurpose: The goals of this study are to compare the mRNA profiles of LAP2 deficient human adipose-derived stem cells (hASCs) with that of control hASCs by NGS-derived RNA-seq to screen differentially expressed genes and possible involved pathways. Methods: Total RNA was isolated from hASCs transfected with shLAP2 and shNC. RNA sequencing of control and LAP2 deficient cells (each with 2 biological replicates) was performed using BGISEQ-500 sequencing system. Differentially expressed genes (DEGs) between control and LAP2 deficient cells were screened using NOISeq method. KEGG pathway enrichment analysis and Gene Ontology analysis of DEGs were subsequently conducted. Results: Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the cell genome. The scatterplot showed that depletion of LAP2 resulted in the upregulation of 106 genes and downregulation of 91 genes. KEGG pathway analysis suggested that LAP2 knockdown led to the change of several important pathways, including cell cycle, MAPK, and NF-B signaling pathways that are critically related to cell growth and differentiation. Conclusions: Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. SOURCE: Yiman Tang (tangyiman085@gmail.com) - Peking University
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