Ecsod

These analyses were performed for MAFbx/Atrogin-1, MuRF1 and Gapdh mRNAs in plantaris muscles as described^{23 (link)}. Real-time PCR was performed using primers for MAFbox/Atrogin 1 and 18S rRNA from Applied Biosystems (Foster City, CA). Immunoblot analysis—Immunoblot analysis was performed as described^{6 (link)} with the following primary antibodies: CuZnSOD (Abcam), MnSOD (Abcam), EcSOD (R&D, Minneapolis), peroxisome proliferator-activated receptor γ coactivator 1-α (PGC- 1α) (Chemicon/Millipore), cytochrome oxidase IV (COX IV) (Invitrogen), cytochrome C (Cyt C) (Cell Signaling, Danvers), malondialdehyde (MDA) (Academy Biomedical Company, Inc., Houston) and 4-hydroxynonenal (4-HNE) (ab48506, Abcam). Hybridoma for antibodies against MHC I (BA-F8), MHC IIa (SC-71) and MHC IIb (BF-F3) were purchased from German Collection of Microorganisms and Cell Cultures. All the bands for analysis have been validated previously^{3 (link), 6 (link), 24 (link)}. OxyBlot Protein Oxidative Detection Kit (Millipore) was used for immunblot detection of carbonylated proteins. Immunoblots were analyzed by Odyssey Infrared Imaging System (LI-COR Biosciences, NE) and quantified by Scion Image software.

Immunoblot analysis was performed as described[24 (link)] with the following primary antibodies: EcSOD (R&D, Minneapolis), VCAM-1. Immunoblots were analyzed by Odyssey Infrared Imaging System (LI-COR Biosciences, NE) and quantified by Scion Image software.