RNA-seq analysis of 6-week-old WT or AKO mice after a 3-day cold challenge

For the RNA-seq analysis of SUB, total RNA was extracted from 6-week-old WT or Adipoq-cre/Nectin2 loxp/loxp (AKO) mice after a 3-day cold challenge using TRIzol reagent in accordance with the manufacturer's protocol. The RNA libraries were sequenced on the illumina NovaseqTM 6000 platform and analyzed by LC Bio Technology Co., Ltd (Hangzhou, China). The sequence data was checked with FastQC (0.10.1) and RSeQC (v4.0.0). Clean data were mapped to GRCm38 with HISAT2 (v2.2.1). Following transcriptome assembly and merging (String Tie, 2.1.6), DEseq2 was employed to quantitatively analyze the transcripts, compare differences, and visualize the results.