Figure 1. Mitochondrial dysfunction in CAV1^−/− cells.

(A) Wt (white bars) and CAV1^−/− MEF (black bars) were cultured with 2-DG. After 48 hours cell number was determined and expressed with respect to the initial number of cells (t₀). (B) Apoptosis, analysed by flow cytometry via binding of annexin V and staining with propidium iodide, promoted by 5mM 2-DG. (C) Apoptosis promoted by DCA, some cells were pre-treated with the antioxidant BHA. (D) Levels of ROS in cells incubated during 24 hours with DCA. The results are expressed as the relative H2DCFDA fluorescence with respect to untreated wt cells. (E) ΔΨ m of CAV1^−/− with respect to wt MEF. (F) Oxygen consumption by wt and CAV1^−/− MEF expressed as the routine flux control ratio. (G) Western blotting analysis of CAV1 (plasma membrane), Rab11 (recycling endosomes), GM130 (Golgi complex), Sec61 (ER) and cytochrome C (Cyt C, mitochondria) in purified wt and CAV1^−/− mitochondria (M), homogenates (H) and in a crude fraction that contains mitochondria and associated ER (cM). (H) Ratios of oxygen consumption in wt (white bars) and CAV1^−/− mitochondria (black bars) purified from mice liver. Statistical significances were determined in at least 5 independent experiments or 10 mice using the Student’s t test, *P<0.05, **P<0.01.