Fig. 2

CEP triggers the accumulation of mitophagosomes in human breast cancer cells. MDA-MB-231 and MCF-7 cells were treated with CEP (4 μM) for 24 h. a Whole cellular lysates (WCL), mitochondrial (Mito) and cytosolic (Cyto) extracts were prepared and subjected to Western blot analysis using antibodies against LC3B-I/LC3B-II. GADPH and COX IV were used as a loading control. The ratio of LC3-II/LC3-I was quantified in three independent experiments (mean ± SD, *P < 0.05 or **P < 0.01). b-c Confocal microscopy images of cells treated without or with CEP (4 μM) for 24 h after co-expressing RFP-mito and EGFP-LC3; scale bars: 10 μm. Quantification of EGFP-LC3 and RFP-mito colocalization puncta in 50 cells of three independent experiments (**P < 0.01). d Representative TEM images of MDA-MB-231/MCF7 cells treated with or without CEP, red arrows indicate mitochondrial. Scale bars: 2 μm