Research Paper Volume 4, Issue 10 pp 664—673

Figure 3. CBS depletion induces mild mitochondrial dysfunction. (A) Left panel: CBS-depleted HUVEC were subjected to high resolution respirometry. The experimental regime started with routine respiration, followed by addition of oligomycin, and stepwise titration of FCCP. Finally, respiration was inhibited by sequential addition of rotenone and antimycin A (Ant.A). Respirometry results were normalized to citrate synthase activity; data are represented as mean ± SE (n=6 and 7 for CBS knockdown and control, respectively). Right panel: Respiratory control ratio was determined as the ratio of uncoupled respiration to oligomycin-inhibited respiration. (B) The electric potential of the inner mitochondrial membrane was measured in situ using flow cytometry in intact cells stained with the JC-1 fluorescent probe. The ratio of cells with high to cells with low mitochondrial membrane potential was calculated as described in the methods. Data are represented as mean ± SE (n=3).