Fig. 2: The mtAP2s are required for the normal morphology and function of the mitochondrion in T. gondii.

a, b Volcano plots highlighting the differential expression genes (DEGs) (a) and the differential expression proteins (b) of ATc versus vehicle or rapamycin versus vehicle-treated in four mtAP2 knockdown lines. The cutoff for DEGs was a p-value < 0.01 and ± 2-fold change, while the cutoff for differential protein expression was a p-value < 0.05 and ± 1.5-fold. X-axis shows log2 fold change, Y-axis shows -log10(P-value). c Electron micrographs of wild-type line and four mtAP2s knockdown lines for 72 h ± ATc. Insets (red) show representative mitochondrion. Scale bar, 500 nm. d, e Extracellular flux (Seahorse) analysis of mtAP2s knockdown lines and ΔmtAP2-1. Delta values of the basal oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) in response to downregulation of four mtAP2s are shown (p-values, top, comparing the -ATc and +ATc condition and the numerical delta values are provided (red), bottom). Bars represent the means of three independent experiments. Dots indicate the means of these experiments, each based on >18 technical replicates. Error bars represent mean ± SD. Statistical significance was tested using a one-way ANOVA, followed by Bonferroni test for multiple comparisons (d). ECAR and OCR traces of a mitochondrial stress test comparing RH, ΔmtAP2-1 and a mutant defective in heme synthesis (Δ coproporphyrinogen III oxidase, CPO/ Δ coproporphyrinogen III dehydrogenase, CPDH)³³. Oligomycin (2.5 µM), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, 2 µM), and antimycin A (0.5 µM) were injected after the 3rd, 6th, and 9th data points, respectively. The displayed traces are from a single representative experiment performed three times independently. Data points show the means and error bars indicate mean ± SD of >12 technical replicates (e). Source data are provided as a Source Data file.