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Each mouse pup was genotyped at postnatal days 2 to 6 by activity gel to detect human Cu/Zn-SOD activity, by single-strain conformational polymorphism (SSCP) to detect the human Cu/Zn-SOD G93A mutant transgene, and by polymerase chain reaction (PCR) to detect the human BCL-2 transgene. Activity gel was performed on hemolysates (40), and SSCP (2) and PCR (14) were performed on purified genomic DNA (QIAamp; Qiagen, Chatsworth, CA) prepared from 200 μl of blood and 1 cm of tail. All genotyping was performed at least twice for each pup. After completion of the genotyping, each pup was given an arbitrary code and, from then on, was assessed for clinical and morphological characteristics by investigators blinded to the genotype; the blind was broken only at the end of the study.
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We thank A. B. Naini and R. Yokoyama for performing the SOD assays, M. Schoenebeck for preparing phrenic nerve sections, M. Gurney for kindly providing the information about the number of copies in the transgenic G93A mice and the G1H mice, and R. E. Burke and C. M. Troy for their insightful comments on the manuscript. This work was supported by the Muscular Dystrophy Association, the National Institute of Neurological Disorders and Stroke (grant NS01724), the Swiss National Foundation (grant 31-4331695), the Lowenstein Foundation, and the Parkinson's Disease Foundation. S.P. is a recipient of the Irving A. Hansen Memorial Foundation Award and the Cotzias Award from the American Parkinson Disease Association
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We thank A. B. Naini and R. Yokoyama for performing the SOD assays, M. Schoenebeck for preparing phrenic nerve sections, M. Gurney for kindly providing the information about the number of copies in the transgenic G93A mice and the G1H mice, and R. E. Burke and C. M. Troy for their insightful comments on the manuscript. This work was supported by the Muscular Dystrophy Association, the National Institute of Neurological Disorders and Stroke (grant NS01724), the Swiss National Foundation (grant 31-4331695), the Lowenstein Foundation, and the Parkinson's Disease Foundation. S.P. is a recipient of the Irving A. Hansen Memorial Foundation Award and the Cotzias Award from the American Parkinson Disease Association.
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