Fig. 4

Depletion of TgANT leads to disruption of mitochondrial morphology. a HA endogenous tags were added to the C terminal of the mitochondrial outer membrane protein TOM40 of the Δant and RHΔku80 strains. Mitochondrial outer membrane morphology was visualized by IFA. Two different mitochondrial phenotypes were identified in Δant, which we named ball-like and broken (yellow arrowhead). GAP45 (green) was used as a marker of the parasite membrane; TOM40 (red) was used as a marker of the parasite mitochondrial outer membrane; DAPI (blue) was used to stain nuclei. Mitochondrial morphology of intracellular parasites was scored as indicated. Scale bar: 2 μm. b Randomly selected vacuoles (n = 100) from two independent experiments were quantified. No significant changes were seen in the RHΔku80 strain, but the mitochondrion of the ant gene knockout parasites showed severe morphological defects, classified as “broken” (prevalence: appox. 27%) and “ball-like” (prevalence: approx. 26%). c Representative transmission electron microscopy images of Δant and RHΔku80 parasites. The deletion of ant resulted in a loss of density and a reduced number of mitochondrial cristae. The mitochondria of RHΔku80 parasites had a regular morphology with a clear cristae structure. M, Mitochondrion. Scale bars: 200 nm