Am10031pu s

Following fixation with 4% paraformaldehyde, the GMSCs were permeabilized with methanol and blocked with 5% BSA in PBS for 20 min. Subsequently, the GMSCs were immunolabeled with antibodies against CD90 (rabbit anti-human monoclonal antibody, 1:100, EPR313, Abcam) S100A4 (rabbit anti-human monoclonal antibody, 1:200, EPR2761(2 (link)), Abcam), vimentin (rabbit anti-human monoclonal antibody, 1:100, EPR3776, Abcam) and cytokeratin (mouse anti-human monoclonal antibody, 1:100, AM10031PU-S, OriGene Technologies, Inc.). Following incubation, the cells were washed with PBS, incubated with fluorescein isothiocyanate-conjugated secondary antibody and counterstained with DAPI. The samples were observed under a fluorescence microscope (Olympus Corporation).

Briefly, endogenous peroxidase activity within the sections was quenched by incubating the sections with 3% H₂O₂ for 10 min following dewaxing and hydration. GMSCs were incubated with primary antibodies against vimentin (rabbit anti-human monoclonal antibody, 1:100, EPR3776, Abcam), CD90 (rabbit anti-human monoclonal antibody, 1:100, EPR313, Abcam), CD73 (rabbit anti-human monoclonal antibody, 1:200, EPR6114, Abcam) and cytokeratin (mouse anti-human monoclonal antibody, 1:100, AM10031PU-S, OriGene Technologies, Inc.) for 30 min at room temperature, followed by incubation with a secondary antibody. In negative controls, the primary antibody was replaced with PBS. The sections were then counterstained with hematoxylin.