FIGURE 7: Analysis of the effect of Erg-GTr and Erg-GTr₂ on the cytoplasmic Ca²⁺ concentration. Stationary phase parasites were loaded with Fura-2AM, acridine orange, or rhodamine 123 to analyze, immediately after addition of Erg-GTr (1.4 μM) or Erg-GTr₂ (2.3 μM), the changes in the cytoplasmic Ca²⁺ concentration, acidocalcisome alkalinization, and mitochondrion membrane potential, respectively.

(A) Relative Ca²⁺ concentration given as normalized OD_340/380 ratio in the presence or absence of extracellular Ca²⁺. Arrows indicate addition of Erg-GTr or Erg-GTr₂.

(B) Effect of ergosterone-triazol hybrid molecules on the alkalinization of acidocalcisomes. The excitation wavelength was 488 nm, and the emission wavelength was 530 nm. Nigericin (2 μM) was used as control to complete organelle alkalinization.

(C) Relative rhodamine 123 fluorescence before and after addition of Erg-GTr or Erg-GTr₂. FCCP (1 μM) was employed as control for total mitochondrion depolarization. Fluorescence was also register at 488 nm excitation and 530 nm emission. All graphics are representative charts of one of at least three experiments that showed essentially the same results.