GSM9325027: Adrb2_cKO_RTX_12H_1; Mus musculus; RNA-Seq
Cells were isolated from Adrb2 cKO C57BL/6J mice in first anagen after 12 hr injection with RTX. Mouse back skin containing the damaged regions was dissected, and the fat layer was removed by scraping with a surgical scalpel. The dermal fraction was isolated by incubating the skin in a collagenase solution in HBSS on an orbital shaker for 45-60 minutes at 37°C with scraping of the dermis. Following collection of the dermis, the remaining skin was incubated in trypsin-EDTA for 25-30 minutes at 37°C to collect the epidermis. Single cell suspensions of each fraction were obtained by filtering through a 70-mm and 40-mm filter. Cell suspensions were then centrifuged for 8 minutes at 350g at 4°C and resuspended in 5% FBS with a CD16/F32 Fc blocking antibody (BioLegend 101302, 1:100). Libraries were constructed according to the 10x Genomics Single Cell v3.1 protocol