Figure 4

Loading of T. torquatus RBC with citosolic Ca²⁺ Fluo-4 and mitochondrial Ca²⁺ Rhod-2. A) T. torquatus RBCs loaded with citosolic Ca²⁺ indicator Fluo 4 AM (Left), mitochondrial Ca²⁺ indicator Rhod 2 AM (center) and merge of Fluo 4 and Rhod 2 (right). B) Increase of mitochondrial Ca²⁺ [Ca²⁺]_mt after increase of [Ca²⁺]_c induced by ATP (50 μM) in T. torquaturs RBCs loaded with both Fluo 4 AM and Rhod-2 AM, in the Ca²⁺ medium. C) In a free Ca²⁺ medium ATP did not induce an increase of [Ca²⁺]_c and in this case we did not observe an increase of [Ca²⁺]_mt D and E) Similar to ATP thapsigargin (5 μM) and ionomycin (10 μM) promote an increase of [Ca²⁺]_c with concomitant increase of mitochondrial Ca²⁺.