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. 2020 Jun 18;21(8):e48882. doi: 10.15252/embr.201948882

Figure 3. Characterization of mitochondrial respiratory complexes and their activity in synaptoneurosomes.

Figure 3

  • A
    Respiration activity of permeabilized synaptoneurosomes (upper panel) as compared to permeabilized HEK293T (lower panel). Oxygen concentration (blue line) and oxygen consumption (red line) measured using O2k Oxygraph are shown.
  • B
    Blue native gel electrophoresis (BN‐PAGE) analysis of mitochondrial respiratory chain complexes from synaptoneurosomes as compared to HEK293T mitochondria.
  • C
    Detection of respiratory chain complexes and supercomplexes in SN and HEK293T mitochondria using BN‐PAGE followed by Western blot using antibodies specific for: complex I (Ndufb8), complex II (Sdha), complex III (Uqcrc2), complex IV (Cox6a), and complex V (Atp5a). Note similar composition of OXPHOS supercomplexes and complexes in synaptoneurosomes as in cellular mitochondria.
  • D
    Selected bands (1‐8) visible in blue native gel were excised and analyzed using mass spectrometry (LC‐MS). Proteomic analysis confirms the presence of protein subunits of different respiratory chain complexes in indicated bands. See Table EV1 for identified peptides.