Fig. 5

NCAM protein expression in hBM-MSCs. NCAM is expressed as transmembrane (NCAM-180 and NCAM-140), membrane-anchored (NCAM-125 and NCAM-120), or secreted (any isoform) protein. Western blotting was performed to identify which NCAM isoforms are expressed at the protein level in hBM-MSCs. In gel electrophoresis, polySia may affect the mobility of NCAM. Parallel samples were thus treated with endosialidase (+Endo) to remove polySia. NCAM was detected with a mixture of two primary antibodies. All of the main NCAM isoforms—NCAM-180, NCAM-140, and NCAM-120—were detected from the whole cell lysates of hBM-MSCs. The NCAM-120 and NCAM-125 isoforms could not be distinguished from one another. The NCAM-expressing neuroblastoma cell line kSK-N-SH served as a positive control. Secondary antibody control confirmed the specificity of the labeling and α-Tubulin served as a loading control. MSC mesenchymal stromal cell, NCAM neural cell adhesion molecule