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Figure 3 | Molecular Neurodegeneration

Figure 3

From: Generation and characterization of transgenic mice expressing mitochondrial targeted red fluorescent protein selectively in neurons: modeling mitochondriopathy in excitotoxicity and amyotrophic lateral sclerosis

Figure 3

Thy1-mitoDsRed2 tg mouse generation and characterization. A. Genomic identification of tg mice. Thy1-mitoDsRed tg mice showed a PCR-amplified 208 bp product that was undetected in non-tg mice. B. Western blot analysis of subcellular fractions of Thy1-mitoDsRed tg mouse brain and spinal cord demonstrating the presence of DsRed protein in mitochondrial-enriched fractions but not in soluble protein fractions. Purified recombinant DsRed was loaded as a positive control. Blots were reprobed with antibody to cofilin to show protein loading. Results were replicated using several tg mice (n = 8). C. Western blot analysis showing the tissue distributions and brain regional distributions of DsRed protein expression in Thy1-mitoDsRed tg mice. Crude extracts were used. DsRed protein was detected at varying levels of expression in most CNS regions but not in liver or kidney (4 of 5 lines). Recombinant DsRed was loaded as a positive control. The nitrocellulose membrane was stained with Ponceau S to show protein loading. Results were replicated using several tg mice (n = 8). D. RT-PCR analysis showing the tissue distributions of DsRed mRNA expression in Thy1-mitoDsRed tg mice. DsRed mRNA was detected in brain but not in skeletal muscle, heart, liver, or kidney. VDAC1 mRNA was used as an internal control. Results were replicated using several tg mice (n = 5). E. RT-PCR analysis showing brain regional distributions of DsRed mRNA expression in Thy1-mitoDsRed tg mice. DsRed protein was detected at varying levels of expression in several different CNS regions. VDAC1 mRNA was an internal control. Results were replicated using several tg mice (n = 5).

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Molecular Neurodegeneration

ISSN: 1750-1326