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. 2014 Nov 20;10(11):e1004815. doi: 10.1371/journal.pgen.1004815

Figure 5. NDUFA10 knockdown slightly reduces CCCP-induced Parkin translocation and mitophagy.

Figure 5

(A) In HeLa cells stably transfected to express YFP-Parkin, before CCCP toxification (0 h) YFP-Parkin (green) has a diffuse cytoplasmic distribution in control (ctrl) siRNA treated cells. Following 4 h CCCP, YFP-Parkin co-localizes with mitochondria labeled with ATP5A immunostaining (red). PINK1 siRNA treatment almost completely abolishes YFP-Parkin translocation. (B) Quantification of YFP-Parkin translocation as in A. (C) Stably transfected HeLa cells expressing YFP-Parkin, before CCCP treatment (0 h, ctrl) have a normal (“High”) mitochondrial content. Following 24 h treatment with CCCP, a high proportion of control cells (ctrl) show complete degradation (“none”) or perinuclear aggregated (“low”) mitochondria, visualized by ATP5A immunostaining (red). PINK1 siRNA treatment almost completely abolishes mitophagy. (D) Quantification of mitochondrial content as in C. Histograms indicate mean ± s.d. of triplicate experiments. n>30 cells per experiment. Scale bar  = 20 µm. * P<0.05, ** P<0.01, *** P<0.001, Student's t-test compared with respective conditions in control siRNA.