Fig. 4.
Reintroduction of MARCH5 diminishes the number of SA-β-Gal-positive cells induced by MARCH5 depletion. MARCH5-R-YFP and MARCH5H43W-R-YFP plasmids, which are resistant against MARCH5 shRNA, were transfected into the cells expressing MARCH5 shRNA on day 2. Two days post-transfection, cells were analyzed for the SA-β-Gal assay and mitochondrial morphology. (A) Quantification of SA-β-Gal-positive cells. Approximately 300 cells were counted in several fields and data represent the average of three independent experiments. ***P<0.001 vs MARCH5 shRNA by Student's t-test. (B) The expression levels of MARCH5-R-YFP and MARCH5H43W-R-YFP were determined by immunoblotting. (C) Mitochondria were visualized with 125 nM MitoTracker Red staining and analyzed by confocal microscopy. Boxed areas are magnified on the right. Scale bars: 10 μm.