Fig. 2.

Localization of trypanosomal TrpRSs. (A Top and Center) Double immunofluorescence analysis of a T. brucei cell line expressing Tb-TrpRS1 carrying the Ty1 tag at its carboxyl terminus under the control of the tetracycline-inducible (+Tet and −Tet) procyclin promoter. The cells were stained for DNA by using DAPI, for a subunit of the ATPase, serving as a mitochondrial marker and with a monoclonal antibody recognizing the Ty1-Tag. (A Bottom) Same as Top and Center, but a cell line expressing carboxyl-terminally Ty1-tagged Tb-TrpRS2 was analyzed. (Scale bars: 10 μm.) (B) Immunoblot analysis of total cellular (T), crude cytosolic (C), and crude mitochondrial extracts (M) for the presence of the Ty1-tagged Tb-TrpRS1 and Tb-TrpRS2, respectively. Elongation factor 1a (EF-1a) served as a cytosolic and α-ketoglutarate dehydrogenase (KDH) as a mitochondrial marker.