Figure 8. Antibodies to the recombinant protein localize TbFUT1 to the mitochondrion.

(A) IFA of wild-type bloodstream form (BSF) trypanosomes after staining with affinity-purified anti-TbFUT1 (αFUT1) MitoTracker and DAPI. Comparable patterns were observed for αFUT1 and MitoTracker, suggesting that TbFUT1 localizes to the mitochondrion. (B, C) TbFUT1 conditional null mutants were grown under permissive (+Tet) and non-permissive (-Tet) conditions for 3 days in the case of BSF (B) or 7 days for PCF (C) and imaged.. In both cases, the tetracycline-inducible TbFUT1 pattern is consistent with a mitochondrial localization. (D) BSF trypanosomes induced to express a C-terminally tagged known Golgi glycosyltransferase (TbGnTI-HA₃) were imaged after staining with αFUT1, anti-HA, and DAPI, as indicated. The merged images of two representative cells suggest no significant co-localization between native TbFUT1 and the Golgi-localized TbGnT1. White boxes: areas magnified 1.5× in the rows below. Scale bars: 3 μm.