Fig. 3
From: Proteomic profiling of neuronal mitochondria reveals modulators of synaptic architecture
Purification of distinct mitochondrial subpopulations and validation of proteomic data. a. Quantitative fluorescent western blots demonstrating relative abundance of the mitochondrial markers COXIV and VDAC1 and the nuclear marker telomerase in whole brain lysate and mitochondrial samples. Mitochondrial samples display little nuclear contamination and enrichment of COXIV and VDAC1. b. Quantification of the beta-tubulin loading control signal. c. Purity of mitochondrial isolates was verified with quantitative enrichment analyses utilising the raw proteomic data. Comparative expression of the synaptic markers Snap25, Synpo, Synj1 and Psd3 indicate significant enrichment (****p<0.0001) of the proteins in synaptic mitochondrial preparations. d-f. Left bar chart displays the proteomic average normalised expression values of proteins in synaptic and non-synaptic mitochondria. Right bar chart demonstrates sample protein expression quantified by fluorescent western blots. Proteomic and sample expression of all proteins (HIBCH, OGDH and citrate synthase) follow the same trend thereby providing validation of the proteomic data. Statistical analyses utilized unpaired two-tailed Student’s t-test, n = 3 (** p=<0.01; *** p=<0.001; **** p=<0.0001)