Fig. 6.

Assessment of mitochondrial proteins and DNA. (A-B) Levels of mitochondrial electron transport chain complex proteins were analyzed by western blot of extracts (samples mixed with sample buffers and at room temperature for 10 min without boiling) from primary cortical neurons by using Abcam's total OXPHOS rodent antibody cocktail, n = 3, normalized to CI, no statistical analysis performed. (C-I) Primary cortical neurons exposed for 24 h to 40 µM chloroquine (CQ). (C-E) Western blot analysis of two TCA cycle enzymes aconitase and citrate synthase. (F) Citrate synthase, a key enzyme in the TCA cycle, activity was measured by biochemical method and UV spectrometry. (G) The specific activity levels were then normalized to citrate synthase protein levels as determined in Fig. 6C, E. (H) Measurement of mtDNA copy number by normalizing mtDNA PCR to nDNA PCR. (I) Measurement of mtDNA damage. *p < 0.05 compared to control, n ≥ 3.