BioProject

Gene expression profiling reveals multiple biological functions of oleacein. We evaluated the effects of oleacein on human adipose-derived stem cells. We performed an untargeted whole-genome transcriptome analysis to explore functionality of oleacein in a adipose stem cell-based tool. Overall design: According to the manufacturer's guide, the RNA was extracted using Isogen kit (Nip-pon Gene Co. Ltd., Japan). Then, RNA quantity and quality were determined using the NanoDrop 2000 spectrophotometer (ThermoScientific, USA). DNA microarray analysis was conducted on control and oleacein treated hASCs samples using the GeneChip WT PLUS Reagent Kit (ThermoFisher Scientific) and GeneChip™ Hybridization, Wash and Stain Kit (ThermoFisher Scientific) following the manufacturer's instructions. In brief, complementary DNA (cDNA) was synthesized from 100 ng of RNA solutions. cRNA was synthesized from in vitro transcription of cDNA and then purified and reverse transcribed. Finally, single-stranded cDNA (ss-cDNA) was synthesized, purified, fragmented, and labeled following the manufacturer's instructions. Cartridge Array Hybridization was performed using the Clariom S array (Human; ThermoFisher Scientific) on the GeneChip™ Fluidics Station (ThermoFisher Scientific). Scanning was performed using GeneChip Scanner (ThermoFisher Scientific). The raw image data obtained after scanning were analyzed using the Transcriptome Analysis Console (TAC) software (ver. 4.0.2, ThermoFisher Scientific). The raw data were normalized following the signal space transformation robust multi-chip analysis (SST-RMA) algorithm. Further, gene-level analysis was performed using the Limma Bioconductor package. For differential expression analysis, a One-Way ANOVA followed by an empirical Bayes correction was performed.ar space) were considered as differentially expressed genes (DEGs). A threshold value of FC >2 (in linear space) and p-value <0.05 (one-way between-subject analysis of variance ANOVA) were set to identify as differentially expressed genes (DEGs) in analysing the comparasions to normal hASCs (undiff). After adipocyte differentiation, DEGs were filtered using a linear fold change (FC) of greater than 1.1 and a p-value less than 0.05 for comparing the OLE+diff and diff groups in h-ASCs.