Transcriptome Analysis of Preadipocytes and Adipocytes

Transcriptome analysis was performed as previously described [3 (link),65 (link)]. Briefly, aliquots of preadipocytes and adipocytes were used for RNA extraction, and mRNA was isolated from 1 µg RNA by poly-dT enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (BioLabs, Heidelberg, Germany). After fragmentation, the samples were subjected to the workflow for strand-specific RNAseq library preparation (Ultra Directional RNA Library Prep II, NEB) and 75 bp single-read sequencing was performed on an Illumina NextSeq500 (Shirley, NY, USA). After sequencing, FastQC was used to perform quality control. Differential expression between preadipocytes and adipocytes was tested with the R package DESeq2 v2.7.R [65 (link),68 (link)]. Selected gene expression data have been published previously [3 (link),65 (link)]. The complete data set is available under GEO accession GSE169514. A fold-change of ≥2 and ≤−2 and Benjamini–Hoch-berg-adjusted p values of ≤0.05 were used as criteria to select differentially expressed genes (DEG).