Figure 8

Mitochondrial outer membrane proteins integrity is essential for mitochondria transformation.

(A,C) Mitochondria were isolated from HeLa-DsRed2-mito cells and treated with trypsin at the indicated concentration (% from mitochondria, w/w) for 10 min on ice. For control, 20% (v/v) FBS was added in addition to trypsin. Following three washes, the mitochondria (300 μg/ml) were added to (A) HepG2 cells or (C) healthy fibroblasts, plated in 96 wells plates and were incubated for 1 h. The cells were washed three times in PBS to remove excess mitochondria and the fluorescence was recorded in a plate reader. Error bars represent ± SEM, n = 3, ^*P < 0.004; ^**P < 0.0002. (B,D) Mitochondria were isolated from HeLa-DsRed2-mito cells and treated with trypsin at a concentration of 2% (w/w), at the indicated time periods on ice. For control, 20% (v/v) FBS was added in addition to trypsin. Following three washes, the mitochondria (300 μg/ml) were added to (B) HepG2 cells or (D) healthy fibroblasts, plated in 96 wells plates and were incubated for 1 h. The cells were washed three times in PBS to remove excess mitochondria and the fluorescence was recorded in a plate reader. The florescence of each treatment was calculated as percentage from control cells and represented as relative transformation. In all the experiments, the fluorescence of the mitochondria alone was tested in a plate reader and was found equal to untreated mitochondria. Error bars represent SEM, n = 3, ^*P < 0.02; ^**P < 0.006.