title: GSM3460572: LC_KO_3 [RMN038]; Mus musculus; RNA-Seq
library_construction_protocol: Mouse skin cells were isolated as described previously (Ginhoux et al., 2007). Briefly, mouse ears were split into dorsal and ventral halves and floated in RPMI-1640 medium (Sigma) containing 1 mg/ml dispase (Invitrogen) for 60 min at 37°C to allow separation of epidermal and dermal sheets. The separated epidermis were then cut into small pieces and incubated in RPMI containing 10% serum, Collagenase IV (0.2mg / ml, Roche) and DNAse I (20000U / ml, Roche) for 1 hr at 37°C. Cells were then passed through 19 G needle and filtered through a 100 µM cell strainer (BD Falcon) to obtain a homogenous cell suspension. Langerhans cells has been sorted based on this phenotype: DAPI- CD45+ Ly6G- CD103- MHCII+ CD24+ EpCAM+. post sort purity has been  chacked and only 99.5 % purity sor or above has been selected for bulk RNA sequencing RNA libraries were prepared for sequencing using standard  protocols