GSM9338174: Visium, SSc4; Homo sapiens; OTHER
Skin biopsies were incubated overnight in 0.4% dispase (Life Technologies) in Hank's Balanced Saline Solution (Gibco) at 4°C. The epidermis and dermis were manually separated. The epidermal layer was digested in 0.25% Trypsin-EDTA (Gibco) with 10U/mL DNase I (Thermo Scientific) for 1 hour at 37°C, quenched with FBS, and filtered through a 70uM strainer. The dermis was minced and digested in 0.2% Collagenase II (Life Technologies) and 0.2% Collagenase V (Sigma) in DMEM for 1.5 hours at 37°C, then filtered through a 70uM strainer. Cells were counted and then combined at a 1:1 ratio. Using the 10X Multiome (ATAC + RNA) workflow, single-nuclei were isolated and captured on a Chromium X instrument, and Illumina sequencing libraries prepared following the manufacturer's protocols (CG000338, CG000365). Libraries were sequenced on an Illumina NovaSeq 6000