BCKDH: The Missing Link in Apicomplexan Mitochondrial Metabolism Is Required for Full Virulence of Toxoplasma gondii and Plasmodium berghei
Figure 6
Acetate complementation of P.berghei BCKDH null mutants.
(A) Following in vitro invasion of normocytes with WT or Pbe1a_ko, parasites were allowed to mature in vitro for 20 h in media supplemented or not with 5 mM acetate. Replication of DNA content was taken as measure of parasite maturation. DNA was labelled using Vybrant DyeCycle Ruby Stain and fluorescence intensity was measured by flow cytometry. Highlighted in green is the ring/parasites degenerated early in their development fraction while trophozoite/schizont stage iRBCs are highlighted in orange. The results of two other biological replicate can be found in Fig. S6. (B) Giemsa-stained blood smears showing development of the different strains within normocytes in medium complemented or not with 5 mM acetate. WT mature from ring to schizonts within normocytes in presence of 5 mM acetate. Pbe1a_ko degenerate rapidly within normocytes in normal culture conditions while complementation with 5 mM acetate rescues partially their viability and maturation. Figure shows the various stages of Pbe1a_ko maturation that can be observed over the in vitro culture period. Parasites were cultured in vitro for the times indicated.