Fitc anti mouse ly 6a e sca 1

Flow cytometry was performed for analysis of cell surface antigen expression. The MSCs at passage one were detached using Accutase (Innovative Technologies Inc., San Diego, CA, USA) and resuspend into PBS at a concentration of 1 × 10⁵ cells per 100 µL. The cells were then incubated with fluorescence-labeled primary antibody or isotype control for 30 min in dark condition. The utilized primary antibodies were: FITC anti-mouse Ly-6A/E Sca-1 (Biolegend), PE anti-mouse CD 90, PE anti-mouse CD44, PE anti-mouse CD 146, FITC anti-mouse CD14, PE anti-mouse CD34 (BD, Franklin Lakes, NJ, USA). These samples were analyzed by Accuri™ C 6 Flow Cytometer (BD).

Etanercept (Sigma, Y0001969) was used at 100 uM in vitro and 1 mg/kg in vivo delivery via intraperitoneal injection in 100 uL saline solution. bFGF (Gibco, #13256-029) was used at 10 ng/mL, rMETRNL (R&D Biosystems) was used at 100 ng/mL in vitro and 0.1 ng/dose in vivo. Antibodies used include the following: APC anti-mouse CD45 (1:200, Biolegend 157605), APC anti-mouse CD11b (1:200, Biolegend, 101211), PE anti-mouse CD106 (1:50, VCAM, Biolegend 105713), FITC anti-mouse Ly6A/E (SCA1, 1:50, Biolegend, 108105), Pacific Blue anti-mouse Ly6C (1:200, Biolegend, 128013), AlexaFluoro 488 anti-mouse CD170 (SiglecF, 1:200, Biolegend 155524), AlexaFluoro 488 anti-mouse Ly6G (1:200, Biolegend 127626), APC anti-mouse F4/80 (1:200, Biolegend 123116), Pacific Blue AnnexinV (Biolegend 640926), PE-Cy7 anti-mouse CD11b (1:200, Biolegend 101216). Mouse Meteorin-like DuoSet ELISA (R&D Systems DY6679).