Differential proteome of bone marrow mesenchymal stem cells from osteoarthritis patients R. Rollín, Ph.D., F. Marco, M.D., Ph.D., E. Camafeita, Ph.D.,

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Differential proteome of bone marrow mesenchymal stem cells from osteoarthritis patients R. Rollín, Ph.D., F. Marco, M.D., Ph.D., E. Camafeita, Ph.D., E. Calvo, Ph.D., L. López-Durán, M.D., Ph.D., J.Á. Jover, M.D., Ph.D., J.A. López, Ph.D., B. Fernández-Gutiérrez, M.D., Ph.D. Osteoarthritis and Cartilage Volume 16, Issue 8, Pages 929-935 (August 2008) DOI: 10.1016/j.joca.2007.12.006 Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 2D-DIGE of MSC proteins. Each individual sample (OA and hip fracture subjects without OA) and a pooled reference sample were labeled with Cy5, Cy3 and Cy2, respectively, mixed, and separated on a 2D-PAGE gel. Gels were scanned and a set each of Cy5 (A), Cy3 (B) and Cy2 (C) images were obtained from each gel. An overlay of three dye scan-images was also obtained (D). Osteoarthritis and Cartilage 2008 16, 929-935DOI: (10.1016/j.joca.2007.12.006) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 (A) 2D silver-stained gels of MSC proteins. Spots for which the quantitative statistical analysis revealed >1.5-fold protein expression change and a Student's t test P value less than 0.05 in OA are shown in circles and numbered according to pI and Mw. The circled protein spots were cut out of this gel and subjected to tryptic digestion followed by MS analysis. (B) DeCyder Software 3D view of differentially expressed protein spots in MSCs from patients with OA. Glucos: glucosidase; Cath B: cathepsin B; Stath: stathmin; Cald: caldesmon; Vim:vimentin; C: hip fracture subjects without OA. Osteoarthritis and Cartilage 2008 16, 929-935DOI: (10.1016/j.joca.2007.12.006) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Graphical representation of the functional classification of proteins differentially expressed in MSCs from patients with OA and hip fracture subjects without OA based on the data in Table I. Osteoarthritis and Cartilage 2008 16, 929-935DOI: (10.1016/j.joca.2007.12.006) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Chemotaxis of MSC induced by PDGF-BB. Cells were seeded in the upper compartment of a Transwell insert, and 1ng/ml of PDGF-BB was added to the upper/lower compartments. The cells were allowed to migrate for 4h at 37°C. The upper and lower compartments were separated by a polycarbonate membrane with 8μm pores. Shown is the mean CI±SD; P values from Mann–Whitney U test less than 0.05 were considered significant and marked by an asterisk (∗). Osteoarthritis and Cartilage 2008 16, 929-935DOI: (10.1016/j.joca.2007.12.006) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions