RNA-sequencing of SVF-derived preadipocytes and adipocytes from inguinal white adipose tissue of male mice treated with vehicle, CGRPa, NPY, NE, or SP
To investigate the effects of sensory neuropeptides on preadipocyte and adipocyte function, we treated cultures with vehicle, calcitonin gene-related peptide-alpha (CGRPa), neuropeptide y (NPY), norepinephrine (NE), or substance P (SP) for 10 hours and collected RNA for RNA-sequencing. 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). Preadipocyte or adipocytes were treated with vehicle, CGRPa, NPY, or SP for 10 hours. 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.