Depletion of PfRieske results in mETC failure: (A) Schematic representation of our experimental setup for the membrane potential and hypersensitivity assays. (B) IC50 analysis reveals hypersensitivity of GlcN-treated parasites (PfRieske-iKD) to the mETC inhibitors atovaquone (ATQ), DSM-265, and proguanil (PRG). (C) IC50 analysis confirms no change in the sensitivity of GlcN-treated parasites (PfRieske-iKD) to drugs used as a negative control for (B) Chloroquine (CQ) and Artemisinin (ART). The graphs in B and C are each representative of n = 2 independent experiments, and the numbers are the mean of the technical triplicates performed in each of the two. (D,E) confocal fluorescent microscopy images of MitoTracker-Deep-Red stained parasites grown in the presence or absence of glucosamine, atovaquone, and proguanil (±GlcN/ATQ/PRG). Combinations that do not affect the membrane potential are shown in (D) as controls, while treatment with atovaquone and proguanil or upon PfRieske depletion and proguanil treatment are shown in (E). Scale bar: 5 µm (F) Quantification of the mitochondrial signal in each of the conditions in (D,E). The data shown are representative of the n = 2 independent experiments. Sheokand PK, Pradhan S, Maclean AE, Mühleip A, Sheiner L. Plasmodium falciparum Mitochondrial Complex III, the Target of Atovaquone, Is Essential for Progression to the Transmissible Sexual Stages. Int J Mol Sci. 2024 25(17):9239. PMID: 39273187