Cd49f fitc

The stem cells were dissociated with trypsin, washed with cold PBS, and then separately stained with immunoglobulin G (IgG) or the following monoclonal antibodies conjugated to PE, PerCP, APC, or FITC: Epcam-PE, CD44-FITC, CD29-FITC, CD49f-FITC, CD73-FITC, CD105-APC, CD90-FITC, CD34-PerCP, CD31-FITC, CD45-FITC, and HLA-DR-FITC (all from eBioscience, USA). Upon being washed with PBS, the labeled cells were resuspended, and at least 10⁵ events were acquired by using a BD Accuri™ C6 flow cytometer (BD, USA).

The culture cells were dissociated with trypsin and washed with cold PBS, then stained with IgG or monoclonal antibodies. The following is the information of antibodies: CD29-FITC, CD49f-FITC, CD73-FITC, CD105-APC, CD90-FITC, CD34-PerCP, CD31-FITC, CD45-FITC, and HLA-DR-FITC (all from eBioscience). Upon being washed with PBS, the cells were resuspended and at least 10⁵ events were acquired by using a BD Accuri™ C6 flow cytometer (BD bioscience).
As for immune cells derived from skin tissues of mice, skin tissues were cut into small pieces and digested with mixed enzymes containing 1 mg/ml collagenase (Sigma), 1 mg/ml hyaluronidase (Sigma) and 0.1 mg/ml DNase I (Roche) in a water bath shaker at 37 °C for 60 min. Then, the cell pellets were filtered and centrifuged, subsequently washed with PBS and resuspended in 1640 medium with 10% FBS, lastly stimulated with phorbol myristate acetate (PMA,100 ng/ml), Ionomycin (1 ug/ml) and Brefeldin A (10 µg/ml) at 37 °C incubator for 5 h. After stimulation, the cells were stained with CD3-FITC (eBioscience) and γδ TCR-PE-Cy7 (Biolegend), and fixed and permeabilized with the Cytofix/Cytoperm™ Kit (BD Biosciences). After that, the cells were stained with IL-17A-PE (BD bioscience). The labeled cells were resuspended and at least 10⁵ events were acquired by using BD Fortessa.