GSM8972806: BAT-DHM_HFD-3; Mus musculus; RNA-Seq
For single-nucleus isolation, tissues were minced into 1-mm pieces in 1 mL of pre-chilled lysate, then homogenized with an additional 2 mL of lysate using a tissue homogenizer (5 slow strokes). After incubation on ice for 5 min, 4 mL of pre-cooled 2% BSA was added to terminate the reaction. Samples were filtered through a 30-μm cell strainer, and the filtrate was centrifuged (300 ×g, 5 min, 4°C). Pellets were resuspended in 5 mL of 2% BSA and centrifuged again under the same conditions. Nuclei were resuspended in 500 μL of resuspension buffer (1× PBS, 2% BSA, 0.1% RNase inhibitor), stained with propidium iodide (PI), and sorted on a BD FACSAria II flow cytometer to exclude debris For snRNA-seq, nuclei were loaded onto a 10x Chromium chip (10X Genomics Chromium Single-Cell 3' Kit v3) targeting 8,000 cells per sample. cDNA amplification and library preparation followed the manufacturer's protocol. Libraries were sequenced on an Illumina NovaSeq 6000 (LC-Bio Technology, China) with paired-end 150-bp reads (minimum depth: 20,000 reads/cell).
GEO
GSM8972806_r1