Fig. 2.

Metabolic changes after NDH2 ablation. (A) Ablation of NDH2 decreases the amount of acetate excreted by the parasites. The amount of acetate, pyruvate and alanine excreted by the WT cells and the Δnubm and/or ndh2 mutant cell lines (given as nmole/h/mg of protein) was determined by ¹H NMR spectrometry as previously described [14]. Three biological replicates were performed for each cell line, except for WT (7 replicates) and Δnubm (8 replicates). The standard deviations for each condition are indicated. Δndh2 clone a3-10 and Δnubm ndh2 cKO clone 4 were used. (B) Characterisation and relative quantification of the total fatty acids present in the lipid extracts of duplicate biological replicates of each of the various clones. This was done by base hydrolysis and subsequent conversion of the fatty acids to their methyl esters allowing analysis by GC–MS as previously described [23].