GSM7090416: visium_sham_male; Mus musculus; OTHER
10X Visium protocol: After removing the renal capsule, the kidneys were bisected in a coronal manner to prepare for 10x Genomics Visium sample preparation. To preserve the high quality of RNA for processing, the fresh tissue was immersed in a bath of 2-methylbutane equilibrated with liquid nitrogen. The tissue samples were then stored at -80°C prior to embedding them in optimal cutting temperature compound (OCT). The remaining kidney tissue was fixed overnight in 10% formalin at room temperature, before being transferred to 70% ethanol for storage at 4°C. The tissue was subsequently embedded in paraffin by the Washington University Musculoskeletal Research Center Core. Frozen kidney samples were embedded in cryomolds using OCT on dry ice. The blocks were then stored at -80°C. For preparation of Visium, the blocks were equilibrated to -18°C, and 10 μm sections were mounted onto the active sequencing areas of the 10× Genomics Visium slides. The slides were stored in airtight containers at -80°C until used for spatial library generation. Hemotoxylin and eosin staining were performed according to the 10× Genomics Visium protocol. Visium libraries were prepared according to 10× Genomics Visium manufacturer's instructions (PN-1000185, Lot No. 155614, Rev D).