GSM8894821: KO2; Mus musculus; RNA-Seq
All mice were weighed, anesthetized with isoflurane, and euthanized by CO2 asphyxiation. The head was decapitated and whole brains were harvested and immediately put on ice in cold Hibernate AB complete media. Thick 1mm coronal sections were made through the area encompassing the mediobasal hypothalamus, identified morphologically under a dissecting microscope. The mediobasal hypothalamus and trace amounts of surrounding regions in the hypothalamus were manually microdissected and dissociated following the protocol in PMID: 17545985, with adjustments as in PMID: 28355573. All tissues and cell solutions were kept on ice whenever possible. The 10x Genomics Chromium Controller was used to capture single cells, with each treatment condition from each donor islet preparation as 1 sample. cDNA libraries were prepared using 10x Genomics Chromium Next GEM Single Cell 3' version 3.1 reagents.