Cd207 apc

Whole back skin was isolated from mice at P4.5 and incubated with 0.5% of Dispase II (Roche, #45147700) at 4°C overnight in 1× Hanks’ balanced salt solution (Sigma-Aldrich, #H2387) without Ca²⁺ and Mg²⁺. Epidermis and dermis were cut into small pieces and digested with Accutase (Gibco, #2310194) containing 0.1% of Liberase DH (Roche, #05401054001) at 37°C and centrifuged at 800 rpm for 20 min. Cells were filtered with a 70-μm nylon mesh and stained with CD140a-Allophycocyanin (APC) (1:150; Invitrogen, #2213095), CD31-APC (1:150; Invitrogen, #219898), CD207-APC (1:150; BioLegend, #B301300), CD117-APC (1:150; Invitrogen, #2162256), α6-integrin-Phycoerythrin (PE) (1:500; Invitrogen, #2196653), and Sca1-Fluorescein isothyocyanate (FITC) (1:300; eBioscience, #E00760-1633) antibodies for 30 min on ice and washed by using 1% fetal bovine serum (FBS), followed by staining with anti–Ephrin-B1 antibodies (1:25; R&D Systems, #BAF473), streptavidin-APC/fire–Alexa Fluor 750 (1:250; BioLegend, #405250), and LEAVE/DEAD Fixable Violet Dead Cell Stain Kit (1:1000; Life Technologies, #L34955) for 30 min on ice intercepted by 1% FBS washing between each staining. Hair matrix keratinocytes were separated from other skin cell populations based on their Ephrin-B1⁺ staining (43 (link)).