Immunolocalisation and expression of keratocan in tendon

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Immunolocalisation and expression of keratocan in tendon S.G. Rees, Ph.D., A.D. Waggett, Ph.D., B.C. Kerr, Ph.D., J. Probert, B.Sc., E.C. Gealy, Ph.D., C.M. Dent, M.D., B. Caterson, Ph.D., C.E. Hughes, Ph.D. Osteoarthritis and Cartilage Volume 17, Issue 2, Pages 276-279 (February 2009) DOI: 10.1016/j.joca.2008.07.007 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Immunolocalisation of keratocan in mature bovine deep digital flexor tendon. (A) Transverse section of compressed tendon immunolabelled with mAb KER-1 (shown in red) which recognises the keratocan core protein and polyclonal antibody 70-XR90 (shown in green) which recognises type I collagen. Areas of co-localisation of keratocan/collagen I immunolabelling are shown in yellow. Cell nuclei are shown in blue. Positive immunolabelling of tendon collagen fibre bundles (solid arrows) and endotenon regions (dashed arrows) is indicated. (B) Transverse sections of compressed tendon (I, II), tensional tendon (III, IV) and cornea (V), immunolabelled with mAb KER-1, and counterstained with DAPI. Images are presented as red/green anaglyphs where positive immunolabelling is shown in green and cell nuclei in red (areas of co-localisation of keratocan immunolabelling with tendon cells are shown in yellow). Positive immunolabelling of tendon collagen fibre bundles (solid arrows) and endotenon regions (dashed arrows) is indicated (I–IV). Tendon sections with the primary antibody omitted showed no immunolabelling (VI). Osteoarthritis and Cartilage 2009 17, 276-279DOI: (10.1016/j.joca.2008.07.007) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Western blot analyses of tissue extracts of cornea, tensional tendon (T-tend), compressed tendon (C-tend) and articular cartilage (Cart), immunolocated with mAb KER-1 which recognises the keratocan core protein. Samples were digested without (A) or with (B) endo-β-galactosidase (+β-endo) to remove KS chains. Loading of samples onto the gels was standardised by loading on the basis of equivalent protein content of tissue extract (30μg). Osteoarthritis and Cartilage 2009 17, 276-279DOI: (10.1016/j.joca.2008.07.007) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions