AmpliSeq Illumina sequencing in male wild-type (Srgn+/+) and serglycin-deficient (Srgn-/-) mice show that serglycin regulates the immune profile of adipose tissue inflammation

Purpose: We here hypothesized that serglycin may have an impact on obese inflammation. To address this, we subjected Srgn+/+ and Srgn-/- male mice to an eight-week high fat/high sucrose diet, followed by an analysis of the effects of serglycin-deficiency on adipose tissue transcriptome. Methods: Ampliseq sequencing was performed on an Ion S5™ XL Sequencer. Total RNA was isolated from eWAT (n=5) using the Direct-zol RNA MiniPrep column kit (The Epigenetics Company, Irvine, CA). cDNA libraries were prepared and amplified with the Ion AmpliSeq™ Transcriptome Mouse Gene Expression Kit (Life Technologies, Carlsbad, CA), following the manufacturer`s instructions. Sequencing was performed on an Ion S5™ XL Sequencer (Thermo Scientific, Waltham, MA). Data had read lengths with average between 98 and 110 bp and high mapping (95% of aligned bases). The expression values were generated by using software Torrent Suite™ (version 5.10.1). Results: The absence of serglycin caused reduced expression of numerous genes linked to inflammatory pathways. Conclusions: This study identifies a role for serglycin, in particular the macrophages, in regulating the immune response in adipose inflammation. mRNA profiles of epidydimal white adipose tissue of male Srgn+/+ (n=5) and Srgn-/- (n=5) mice fed a high sucrose/high fat diet for eight weeks.