Found at: doi:10.1371/journal.pone.0001609.s001 (2.89 MB TIF) Figure S2 RNF5 expression in human myopathies. RNF5 immunostaining of muscle sections from Duchenne Muscular Dystrophy (DMD) and Becker Muscular Dystrophy (BMD) patients compared to a normal muscle cross-section. Arrowheads point to RNF5 staining. Found at: doi:10.1371/journal.pone.0001609.s002 (4.86 MB TIF) Figure S3 RNF5 overexpression is not associated with alter- ations in the dystrophin-glycoprotein complex. Cross-sections of vastus lateralis from treated DTg or control mice were subjected to immunostaining using dystrophin (A) laminin a3 (B) and alpha- dystroglycan (C), revealing a normal pattern of sarcolemmal and extracellular matrix staining. Found at: doi:10.1371/journal.pone.0001609.s003 (4.51 MB TIF) Figure S4 RNF5 overexpression does not prevent proper differentiation and sarcomeric organization of primary myoblasts. Primary myoblasts were isolated from 1 month old animals (RNF5 STg and rtTA RNF5 DTg) and grown in the presence of doxycyclin (2 mg/ml). Differentiation was achieved by switching cells to differentiation medium supplemented with ITS (Insulin Transferrin Selenium supplement). Induction of RNF5 in primary myoblasts prior to differentiation was confirmed by co-immuno- staining with Pax7 (A) and the efficiency of myoblast differenti- ation was monitored by analysis of phase contrast images (bottom References 1. Boonyarom O, Inui K (2006) Atrophy and hypertrophy of skeletal muscles: structural and functional aspects. Acta Physiol (Oxf) 188: 77–89. 2. Glass DJ (2003) Signalling pathways that mediate skeletal muscle hypertrophy and atrophy. Nat Cell Biol 5: 87–90. 3. Tisdale MJ (1997) Biology of cachexia. J Natl Cancer Inst 89: 1763–1773. 4. Jackman RW, Kandarian SC (2004) The molecular basis of skeletal muscle atrophy. Am J Physiol Cell Physiol 287: C834–843. 5. 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Gene Ther 5: 1213–1220. PLoS ONE | www.plosone.org 14 February 2008 | Volume 3 | Issue 2 | e1609