GSM5089673: WT unwounded and wounded skin-CITE-seq_RUN1; Mus musculus; RNA-Seq
Unwounded or a ring with 0.25 mm width of skin adjacent to the wound edge was excised and digested in Liberase TL (Sigma) at 37C for 2.5 hours.  Digests were stopped by places samples on ice and adding EDTA and FBS. Digested tissues were mechanically disrupted by syringe plunger and then filtered through a 70-micron filter to obtain a single cell suspension. Single cell suspensions were stained with anti-CD16/32 before staining with surface fluorescent conjugated antibodies and with surface epitope marking oligo-tagged antibodies and sample specific oligo-tagged Totalseq-A antibodies at predetermined concentrations in a 100 μl staining buffer (PBS containing 5%FBS and 1% HEPES) per 10 million cells. Stained cells were re-suspended in DAPI in FACS buffer prior to cell sorting on a BD FACS Aria II. Hashed samples were pooled at Ctrl,1: D3,1.5, D5:1 ratio prior to scRNA-seq 10X Genomics droplet capture according to manufacturer's protocol.