RNA-sequencing of SVF-derived preadipocytes from inguinal adipose tissue of male mice differentiating under control conditions or treated with CGRP

To investigate the effects of CGRP on adipocyte differentiation, we treated stromal vascular fraction (SVF)-derived preadipocytes from inguinal white adipose tissue (iWAT) of male mice with CGRP during differentiation. RNA-sequencing was performed across a full time course of normal differentiation to identify transcriptional changes induced by CGRP treatment compared to control conditions. Overall design: RNA sequencing analysis of stromal vascular fraction (SVF)-derived preadipocytes from inguinal white adipose tissue (iWAT) of male mice (n = 3 per group, C57BL/6N) undergoing differentiation under control conditions or with CGRP treatment. Total RNA was extracted using TRI reagent (Sigma-Aldrich, T9424) according to the manufacturer's instructions. RNA was further purified, and DNase treated (Zymo, R1013), and integrity confirmed prior to sequencing (Agilent Technologies, 2100 Bioanalyzer), with an RNA integrity number (RIN) greater than 7 as a threshold. Samples were purified by poly-A enrichment before library preparation using the Illumina NovaSeq X Plus Series platform with a paired-end 150 base pair strategy.