GSM8097072: S2, Human, Alcohol-associated Cirrhosis; Homo sapiens; RNA-Seq
Nuclear isolation as described (Filliol et, Nature 2022 Oct;610(7931):356-365). In brief, frozen liver tissue was minced with scissors in 1 ml TST buffer in the well of a 6-well plate for 10 min on ice. The homogenised solution was then passed through a 40 μm cell strainer. An additional 1 mL of TST buffer and 3 ml of 1xST buffer were used to wash the well and passed through a 40 μm cell strainer. The resulting 5 ml of nuclei suspension was centrifuged for 5 min at 500 g at 4°C. The supernatant was discarded, and the pellet resuspended in 1 ml of 1xST buffer. The nuclei suspension was then passed through a 35 μm filter. Single-cell RNA-seq libraries were prepared using the Chromium Single Cell 5′ v2 Reagent Kit (10x Genomics) according to manufacturer's instructions, 12 cycles of cDNA amplification and 12 cycles of library amplification were performed, and samples were sequenced on an Illumina NovaSeq 6000 sequencing system instrument at the JP Sulzberger Columbia Genome Center.