Single-cell RNA-sequencing of Gli1-positive cells from different compartments of the adult mouse epidermis
Single-cell RNA-sequencing of Gli1-positive cells from different compartments of the adult mouse epidermis
Gli1cre/Tom mice were treated with 2 x 3mg tamoxifen i.p. at 7 weeks of age. One week later, full epidermal cells were isolated as described previously (Jaks et al., Nat Genetics. 2008). In brief, dorsal skin was harvested, dermal and adipose tissue was removed, the tissue was cut into strips and subsequently digested in trypsin. Epidermal cells were scraped off the dermis and a single-cell suspension was obtained by magnetic stirring.  The cells were stained with Sca1 and Cd34 antibodies and life-sorted into a fraction of traced cells from the interfollicular epidermis (Gli1/Tomato+ / Sca1+) and a fraction of cells from the hair follicle (Gli1/Tomato+ / Sca1-) using a FACSAria system.   Single-cells from both the hair follicle and interfollicular epidermis fractions were captured separately on Fluidigm C1 Autoprep microfluidic systems. Cell lysis and cDNA synthesis were subsequently performed using a custom protocol (STRT-C1) as described previously (Islam et al., Nat Methods. 2014).  Library preparation from single-cell cDNA was performed using a tagmentation-based approach as described in detail in Islam et al., 2014.
European Bioinformatics Institute
E-MTAB-6031:1772099250-H08_s