Figure 3. Targeting Becn1 alters SOD1 aggregation in ALS transgenic mice. (A) SOD1 aggregation was determined in spinal cord protein extracts derived from Becn1+/+ SOD1G86R and Becn1+/− SOD1G86R mice at the symptomatic stage using western blot analysis. Each well represents an independent animal ordered by crescent genotype and crescent life span. Of note SOD1 oligomers and high molecular (HMW) aggregates are detected in this analysis. “*” represents the same sample from the first lane of Becn1+/+ SOD1G86R mice presented in the left western blot used as a normalization marker to select equivalent film exposure for proper comparison. (B) Mutant SOD1 oligomers were quantified from experiments presented in (A). For the analysis, the average signal of control transgenic mice (*) was normalized to 1 to compare both gels. (C) In parallel, Sod1 mRNA levels were measured by real-time PCR in samples from presymptomatic and end-of- disease stages (right panel). Actin levels were monitored for normalization of gene expression. In (B) and (C), mean and standard error are presented for the analysis of independent animals (at least n = 6 per group). P values were calculated with the Student t test. n.s., nonsignificant differences.