GSM8869544: multiplexed sample set 2, eWAT, HTO; Mus musculus; OTHER
Adipose sNuc-seq was performed as previously described (PMID: 38416649). Briefly, for each 10x run, 8-12 adipose samples were separately homogenized in TST buffer using a gentleMACS Dissociator (Miltenyi Biotec). Lysate was filtered through 40 µm and 20 µm nylon filters (CellTreat), centrifuging and resuspending in fresh buffer each time. After the second wash, each sample was incubated with NucBlue (Thermofisher scientific) as well as an individual hashtag antibody (BioLegend) for 45 minutes. After incubation, nuclei were washed and flow sorted together into RT Reagent B (10x Genomics) using a Beckman Coulter MoFlo Astrios EQ with a 70 μm nozzle to remove poor quality nuclei and count the same number of nuclei per sample. Sorted nuclei were immediately loaded on the 10x Chromium controller (10x Genomics) according to the manufacturer's protocol. Single Cell 3' v3.1 chemistry was used to process all samples and cDNA and gene expression libraries were generated according to the manufacturer's instructions (10x Genomics). Gene expression libraries were multiplexed and sequenced on the Nextseq 500 (Illumina).