GSM9610450: DHT+Ab; Mus musculus; RNA-Seq
Dorsal skin was collected from male C3H/HeN mice after treatment in the vehicle, DHT, or DHT plus anti-DKK3 antibody groups. Hair was removed prior to skin harvest, and excised skin was trimmed to remove excess fat and connective tissue, minced, and dissociated into single cells. Tissue fragments were digested in HBSS containing calcium with Liberase TL (0.5 mg/mL), hyaluronidase (0.1 mg/mL), and DNase I (0.1 mg/mL) for 1.5 h at 37°C with shaking, followed by sequential filtration through 70 μm and 40 μm strainers. Red blood cells were removed using ACK lysis buffer, and debris was depleted by Percoll density-gradient centrifugation. The resulting live single-cell suspension was used for library preparation. Single-cell suspensions were processed using the Chromium GEM-X Single Cell 3' RNA library v4 kit (10x Genomics) according to the manufacturer's instructions. Single cells were partitioned into GEMs with barcoded gel beads, and polyadenylated RNA was captured with barcoded oligonucleotides for reverse transcription and cDNA synthesis. Amplified cDNA was used to generate 3' gene expression libraries, which were sequenced on an Illumina platform.