Spatial transcriptomic profiling of quadriceps muscles from mice fed either standard diet or high-fat diet for 20 weeks
Spatial transcriptomic profiling of quadriceps muscles from mice fed either standard diet or high-fat diet for 20 weeks
Seven-month-old male C57BL/6J mice were fed either a low fat diet (LFD; 10 kcal% fat; D12450J, Research Diets) or a high fat diet (HFD; 60 kcal% fat; D12492i, Research Diets) for 20 weeks. At study termination, mice were euthanized and quadriceps muscles were dissected and immediately flash-frozen in liquid nitrogen. Fresh-frozen quadriceps were cryosectioned into 10 µm cross-sections at the Helmholtz Munich Core Facility Pathology and Tissue Analytics and placed on 10x Genomics Visium spatial gene expression slides. Sections were fixed and stained with DAPI, rat anti-Laminin (1:200, ab11576, Abcam), and rabbit anti-Perilipin (1:100, #57117SF, Cell Signaling), detected with donkey anti-rabbit AlexaFluor555 (1:100, A31572, Invitrogen) and goat anti-rat AlexaFluor647 (1:100, A21247, Invitrogen). Whole-section fluorescence images were acquired on a Zeiss AxioScan 1 slide scanner at 20x magnification. Visium Spatial Gene Expression Slide & Reagent Kit, 4 rxns, PN-1000187 10x Genomics Visium Spatial Gene Expression 3′ barcoded cDNA library-construction workflow for fresh-frozen tissue