Figure 4. Schematic diagram of MAM isolation and purity confirmation of the MAM fraction by WB and enrichment analysis. (A) MAM was isolated from ZDF rat brains by applying differential centrifugation and self-forming Percoll gradient centrifugation. Other cell organelles, such as crude mitochondria, pure mitochondria, and ER were similarly obtained following the multiple centrifuge steps. (B) Western blot analysis of organelle markers in isolated MAM from the brain were enriched for FACL4 and KDEL, and were free from tubulin and cytochrome-C contamination. H: homogenate; Mp: pure mitochondria; Mc: crude mitochondria; ER: endoplasmic reticulum; MAM: mitochondria-associated ER membrane; C: cytosol. DAVID Gene Ontology enrichment analysis of MS-identified proteins. The most enriched (top 10) biological pathways (C), cellular components (D), and molecular functions (E) of the ranked protein list. (F) KEGG pathway enrichment analysis of MS-identified proteins.