Fig. 3
Exaggerated autophagy dysfunction increases SOD1 aggregate accumulation in PACER/SOD1G93A mice. A–C Levels of autophagy-related proteins and SOD1 aggregates were determined in the lumbar spinal cord of symptomatic (P121) SOD1G93A-Tg (n = 5) and PACER/SOD1G93A-Tg (n = 5) mice. Levels of the same proteins in non-Tg (n = 2) and PACER-Tg (n = 2) serve as references. A Western blot analysis of autophagy markers Beclin1, p62 and LC3-II, as well as SOD1 under reduced (+DTT) and non-reduced (−DTT) conditions to detect SOD1 aggregates. β-Actin was detected as a loading control. B Quantification of the fold change of autophagy protein levels as well as total SOD1 (+DTT) in the spinal cord, normalized to β-Actin levels, are shown. C Quantification of SOD1 high molecular weight (HMW) aggregates, as well as Dimers and Monomers under non-reducing (−DTT) conditions was performed using β-Actin levels as a loading control. In B and C statistical analysis using Mann–Whitney U-test was performed. Data is presented as means ± S.E.M. p value: p > 0.05: n.s., non-significant, *p < 0.05; **, p < 0.01