Open Access Research article

Transglutaminase 2 cross-linking activity is linked to invadopodia formation and cartilage breakdown in arthritis

Annie Lauzier, Martine Charbonneau, Marilène Paquette, Kelly Harper and Claire M Dubois*

Author Affiliations

Immunology Division, Department of Pediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, 3001 12th North Avenue, Sherbrooke, Quebec, J1H 5N4, Canada

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Arthritis Research & Therapy 2012, 14:R159  doi:10.1186/ar3899

Published: 4 July 2012

Additional files

Additional file 1:

Figure S1. TG2 expression is increased in synovial tissues in CIA. (A) Representative images (40× magnification) of synovial membrane/cartilage from articular tissue sections of control and arthritic rats at different times (18, 28, and 32 days) after immunization with type II collagen. Tissues were immunostained with TG2. (B, C) Graphs show relative labeling intensity calculated from six random fields for each tissue section in the (B) synovial membrane or in the (C) cartilage. Each column represents mean value ± SEM of total fields from two different tissue sections per individual rat (Ctl, n = 4; CIA, n = 8).

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Additional file 2:

Figure S2. TG2 is linked to the cross-linking activity in synovial tissues. (A) Synovial tissues from control (PBS) and CIA rats were incubated with biotin-pentylamide (PAB) in the presence or absence of 250 μM KCC-009 (KCC) or 100 μM Z-DON and revealed by Western blotting. (B) PAB was injected in PBS or CIA articulations with or without Z-DON (100 μM) and revealed by immunochemistry with streptavidin-peroxidase. Neg Ctl, control tissue without biotin-pentylamide.

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Additional file 3:

Figure S3. Evaluation of CIA progression by using a modified Mankin scoring system. Histopathologic classification of the severity of arthritic lesions was evaluated by using a modified Mankin scoring system. Graph shows mean values ± SEM for control and arthritic rats at different times (18, 28, and 32 days) after immunization with type II collagen (Ctl, n = 4; CIA, n = 8).

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Additional file 4:

Figure S4. Modulation of TGase activity is associated with TG2 regulation. (A-C) TGase activity was measured by using in situ 5-(biotinamido)-pentylamine incorporation assay in (A) C-FLSs or A-FLSs cultured on gelatin matrix. (B) A-FLSs were transfected with control- or TG2-shRNA-expressing lentivirus. (C) C-FLSs were incubated with TGF-β (20 ng/ml), DTT (1 mM), or cystamine (100 μΜ).

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Additional file 5:

Figure S5. Src is implicated in TG2-mediated invadopodia formation. C-FLSs were transfected with a TG2-expressing or a control (empty) plasmid and cultured on gelatin in the presence or absence of PP2. After 24 hours, the percentage of invadopodia-forming cells was counted. Values are expressed as the mean ± SEM (*P < 0.05; **P < 0.01; ***P < 0.001).

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