R. J. U. Lories, M. D. , Ph. D. , J. Peeters, B. Sc. , K. Szlufcik, Ph

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Deletion of frizzled-related protein reduces voluntary running exercise performance in mice R.J.U. Lories, M.D., Ph.D., J. Peeters, B.Sc., K. Szlufcik, Ph.D., P. Hespel, Ph.D., F.P. Luyten, M.D., Ph.D. Osteoarthritis and Cartilage Volume 17, Issue 3, Pages 390-396 (March 2009) DOI: 10.1016/j.joca.2008.07.018 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Running wheel exercise performance in Frzb−/− and wild-type mice. (A) Median daily distance ran at the age of 6 months in male and female mice. (B) Median daily distance ran at the age of 1 year in male and female mice. (C) Mean monthly distance ran in male Frzb−/− and wild-type mice. (D) Mean monthly distance ran in female Frzb−/− and wild-type mice (*indicates P<0.05 in Frzb−/− vs wild-type mice; #indicates P<0.05 in male vs female mice; n=10–13 mice at 6 months and n=5–7 at 1 year). Osteoarthritis and Cartilage 2009 17, 390-396DOI: (10.1016/j.joca.2008.07.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Histomorphology of the knee joint in a wild-type mouse after 1 year of free running. (A) Overview of the knee anatomy on the coronal section. Patella, femur and tibia are labeled. Arrows indicated zones of synovial hyperplasia. Two boxes highlight zones presented more in detail in (B) and (C) (magnification 25×, safranin O/hematoxylin staining). In (B) damage to the superficial layer and loss of proteoglycan staining in the deeper layer of the articular cartilage is shown. Arrows indicate zones of proteoglycan loss, white arrowheads point toward synovitis and black arrowheads show synovial fibrosis. (C) Osteophyte formation indicated by arrowheads. Arrows indicate synovitis. Osteoarthritis and Cartilage 2009 17, 390-396DOI: (10.1016/j.joca.2008.07.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 X-ray analysis of osteoarthritis in running and control mice. (A) Representative images of the different scores used. All images are from wild-type mice. No specific features were found in Frzb−/− mice. (B) X-ray scores in running and control male mice (n=10 control mice in each group; n=4 Frzb−/− and 6 wild-type mice in the running group; both knees were scored for each mouse). (C) X-ray scores in running and control female mice (n=9 Frzb−/− and 10 wild-type control mice; n=7 Frzb−/− and 4 wild-type mice in the running group; both knees were scored for each mouse). Osteoarthritis and Cartilage 2009 17, 390-396DOI: (10.1016/j.joca.2008.07.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histomorphological signs of osteoarthritis in Frzb−/− and wild-type mice. (A) Cartilage damage scores and (B and C) synovitis scores in males (B) and females (C) after 1 year of running exercise. Data are shown as box-plots with medians, quartile and percentile 10–90 (n=12 vs 12 knees from 6 males per group and 8 vs 14 knees from 4 wild-type and 7 Frzb−/− females). Osteoarthritis and Cartilage 2009 17, 390-396DOI: (10.1016/j.joca.2008.07.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Analysis of muscle fiber composition in Frzb−/− and wild-type mice. (A) Immunofluorescent staining for type I (green) and type IIa (blue) fibers in the soleus muscle of wild-type mice and isotype control. (B) Immunofluorescent staining for type IIa (blue) and type IIb (background darkblue) in the extensor digitorum longus muscle of wild-type mice and isotype controls. (C) Muscle fiber surface in Frzb−/− and wild-type male and female mice. (D) Number of muscle fibers in Frzb−/− and wild-type male and female mice (individual data points are shown; dark squares represent Frzb−/− and open circles represent wild-type mice; n=3 vs 3 and 3 vs 2 for male mice and 4 vs 3 for female mice; per mouse muscles from the right and left paw were analyzed). Osteoarthritis and Cartilage 2009 17, 390-396DOI: (10.1016/j.joca.2008.07.018) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions