Expression and function of the insulin receptor in normal and osteoarthritic human chondrocytes: modulation of anabolic gene expression, glucose transport.

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Expression and function of the insulin receptor in normal and osteoarthritic human chondrocytes: modulation of anabolic gene expression, glucose transport and GLUT-1 content by insulin S.C. Rosa, A.T. Rufino, F. Judas, C. Tenreiro, M.C. Lopes, A.F. Mendes Osteoarthritis and Cartilage Volume 19, Issue 6, Pages 719-727 (June 2011) DOI: 10.1016/j.joca.2011.02.004 Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 In situ Immunofluorescence staining of the InsR in normal human cartilage. (A) representative image of the InsRbeta subunit (green) in normal human cartilage sections (N=3) counterstained with DAPI (blue) to allow nuclei visualization, viewed with a 200× magnification (inset: 400× magnification). (B) representative image of the negative control (N=3) viewed with a 200× magnification (inset: 400× magnification). Osteoarthritis and Cartilage 2011 19, 719-727DOI: (10.1016/j.joca.2011.02.004) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 InsR protein levels in chondrocytes from normal and OA human articular cartilage. (A) preInsR and InsRbeta protein content in total cell extracts of chondrocyte cultures established from normal (N=6) and OA (N=6) non-pooled cartilage samples. The image shown is representative of the results obtained in normal and OA chondrocytes. MW=molecular weight marker; N=normal chondrocytes; OA=osteoarthritic chondrocytes. (B) Immunofluorescence staining of the InsRbeta subunit (green) expressed in normal and OA chondrocytes counterstained with DAPI (blue) to show the cell nuclei, as described under “Immunofluorescence staining” in the Materials and methods section. The images shown are representative of the results obtained in 3 normal and 3 OA independent chondrocyte cultures; NC=Negative control. Osteoarthritis and Cartilage 2011 19, 719-727DOI: (10.1016/j.joca.2011.02.004) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Evaluation of the functional state of the InsR expressed in normal and OA human chondrocytes. (A) InsRbeta phosphorylation in chondrocyte extracts from 1 normal and 2 OA cartilage samples untreated (Control) or treated with insulin 10nM for the indicated periods. Akt phosphorylation in total cell extracts from 4 normal (B) and 4 OA (C) non-pooled independent chondrocyte cultures treated with 1 or 10nM insulin for 30min relative to untreated cells. Osteoarthritis and Cartilage 2011 19, 719-727DOI: (10.1016/j.joca.2011.02.004) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Regulation of GLUT-1 content by insulin in normal and OA chondrocytes. Total GLUT-1 protein levels in (A) normal (N=6) and (B) OA (N=6) chondrocytes treated with 1 or 10nM insulin for 48h relative to untreated cells. Osteoarthritis and Cartilage 2011 19, 719-727DOI: (10.1016/j.joca.2011.02.004) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions