Fig. 1: Analysis of transgenic M. smegmatis expressing trypanosomal ASCT.

a ASCT activity was measured spectrophotometrically in 50 mM Tris–HCl (pH 7.4) at 37 °C using the mycobacterial cell lysates. Reactions were initiated by the addition of succinate, and progress was monitored by following the production of 5-thio-2-nitrobenzoic acid, as detected by absorbance at 412 nm. Data represent the specific activity of each strain and are expressed as mean ± SD from three independent experiments. Activities were compared among lysates from M. smegmatis expressing TcASCT or TbASCT, as well as from a control strain harbouring the empty plasmid (pYUB28b). Comparisons were performed using two-tailed one-way ANOVA with post hoc Tukey’s multiple comparisons tests, as indicated. ****p < 0.0001. b Overexpression of trypanosomal ASCT was confirmed by western blotting analysis using anti-ASCT as the primary antibody. Proteins corresponding to ASCT in the lysates ran as bands of sizes similar to those of the purified recombinant trypanosomal ASCTs. Lanes: M marker; 1: M. smegmatis TcASCT lysate; 2: recombinant TcASCT; 3: M. smegmatis TbASCT lysate; 4: recombinant TbASCT. Molecular weights (kDa) of the marker ladder are indicated to the left. Unedited Western blot picture is provided in Supplementary Data 1.