Figure 3. FTVII enforces expression of sLe^X predominantly on N-linked glycoproteins in both BM-hMSCs and A-hMSCs.

(A): Representative flow cytometry histograms of HECA452 mAb and anti-CD44 mAb staining in hMSCs that were untreated (grey line), FTVII-treated (bold line), treated with the protease bromelain (dotted line) or FTVII-treated followed by bromelain treatment (dashed line). Grey filled histogram represents staining with isotype control. (B): HECA452 and CD44 western blot analysis of untreated and FTVII-treated hMSCs +/− bromelain-treatment. (C): HECA452 Western blot analysis of untreated and FTVII-treated hMSCs +/− sialidase-treatment. Lysates of KG1a cells serve as a positive control. (D): HECA452 and CD44 western blot analysis of untreated and FTVII-treated A-hMSCs +/− N-glycosidase F-treatment, indicating that CD44 displays sLe^X on N-glycans. Lysates of KG1a cells serve as positive control. For all figures, data are representative of experiments performed on both A-hMSCs and BM-hMSC cultures derived from at least ten individuals. Abbreviations: FTVII, Fucosyltransferase VII; BM-hMSCs, Bone Marrow-derived human Mesenchymal Stem Cells; A-hMSCs, Adipose-derived human Mesenchymal Stem Cells.