Anti human integrin αvβ3 pe

Cells were collected in PBS and stained with anti-human integrin αvβ3 PE (1:100, Abcam, ab7166, Cambridge, U.K.) at 4°C for 30 min. To examine the percentage of integrin αvβ3 in patients’ samples, 1 mg/ml collagenase (Sigma–Aldrich, San Francisco, U.S.A.), 2 units/ml hyaluronidase (Sigma–Aldrich, San Francisco, U.S.A.), and 0.1 mg/ml DNase (Sigma–Aldrich, San Francisco, U.S.A.) were used to digest the tissues into single cells, the anti-human CD45 APC (1:100, eBioscience, CA, U.S.A.) and anti-human integrin αvβ3 PE (1:100, Abcam, Cambridge, U.K.) were stained at 4°C for 30 min. After washing, the data were collected by BD Canto II (BD Biosciences, NY, U.S.A.). 7-AAD was used to exclude the dead cells. IgG (1:1000, Abcam, Cambridge, U.K.) was used as the negative control. For integrin αvβ3⁺ cell sorting, BD FACSARIA III was used to sort PE positive cells.