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Figure 3


Fig. 3. TbDLP shows a dual localization. A transgenic T. brucei cell line expressing C-terminally HA-tagged TbDLP was analyzed by immunofluorescence. HA-tagged TbDLP was detected using anti HAtag antiserum Y-11 (Santa Cruz Biotechnology) (red). Nuclear (N) and kDNA (K) were visualized by DAPI-staining (blue). The images show 3D-reconstructions from optical sections obtained by confocal microscopy. (A) Co-staining of HA-tagged TbDLP (red) with an antiserum recognizing Hsp60 (green), a marker protein for the mitochondrial matrix. (B) Same as A, but only the nucleus-kDNA region is shown and co-staining was done with the FP marker tomato lectin (TLect) (green). (C) Same as B, but co-staining was done with the antibody YL1/2 (green), which recognizes tyrosinated {alpha}-tubulin and serves as a marker for the basal body (BB) (Sherwin et al., 1987). (D) Same as B, but co-staining was done with the monoclonal mouse antibody L3B2 (shown in green), which recognizes the flagellar attachment zone (FAZ) (Kohl et al., 1999). (E) Schematic drawing of the nuclear-kDNA regions of T. brucei. The localization of the fraction of TbDLP that is required for endocytosis is indicated relative to intracellular structures and organelles. CM, cell membrane; FL, flagellum; FP, flagellar pocket; MT, mitochondrion. Bars, 1 µm.





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