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. 2010 Dec 27;191(7):1395–1411. doi: 10.1083/jcb.201006098

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© 2010 Paul et al.

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Figure 5. — TRAF6 is required for the activation of the ubiquitin–proteasome and autophagy systems in denervated skeletal muscle. 3-mo-old TRAF6^f/f and TRAF6^mko mice were subjected to denervation for 10 d, and TA muscles were isolated for biochemical analyses. (A) Transcript levels of MAFbx and MuRF1 (measured by QRT-PCR assays) were significantly lower in denervated TA muscle of TRAF6^mko mice compared with TRAF6^f/f mice. (B) The expression levels of autophagy-related genes LC3B, Beclin1, and Gabarapl1 were also significantly reduced in denervated TA muscle of TRAF6^mko mice compared with TRAF6^f/f mice. Error bars represent SD. *, P < 0.05 (values significantly different from those of denervated TA muscle of TRAF6^f/f mice). (C) Representative immunoblots presented here demonstrate reduced protein levels of MuRF1, LC3B, and Beclin1 in denervated TA muscle of TRAF6^mko mice compared with TRAF6^f/f mice. Black lines indicate that intervening lanes have been spliced out. (D) Analyses of longitudinal sections of control and denervated TA muscle of TRAF6^f/f and TRAF6^mko mice using transmission electron microscopy. Black arrows point to subsarcolemmal mitochondrial distribution, blue arrows point to intermyofibrillar mitochondria, white arrows point to autophagosomes, pink arrows point to autophagic vacuoles, and red arrows point to mitochondria being engulfed by autophagosome. Bar, 1 µm.