Figure 1

Immunofluorescence and Immuno-electron microscope labelling of the T. brucei TAC. A-F. Immunofluorescence on wild-type cytoskeletons. A. DAPI staining (blue) and Mab 22 labelling (green). B. A merged phase contrast, DAPI staining and Mab 22 labelling of the cytoskeleton in A. The arrows in A and B show the Mab 22 flagellar pocket collar label observed when this monoclonal was initially made and presumably had a very high titre. The arrowhead of A and B indicate the TAC exclusion zone label of Mab 22. C-D. A double labelling immunofluorescence micrograph illustrating a cytoskeleton showing Mab 22 and YL1/2 label. C. DAPI staining (blue) and Mab 22 labelling (green). D. DAPI staining (blue) and anti-basal body staining using YL1/2 (red). E. Merged of C and D. F. Phase contrast merged of E. The arrow in F illustrates the basal body labelling of YL1/2 and the arrowhead indicates the TAC exclusion zone label. In A-F scale bar is 5 μm. G-J. Electron micrographs of longitudinal sections of extracted and Mab 22 probed cytoskeletons. In all images the section traverse a basal and/or pro-basal bodies (BB, pBB). The immunolabelling is clearly observed between the basal or probasal bodies and the outer mitochondrion membrane (black arrow). Kinetoplast is marked as (K) and mitochondrion matrix marked as (M). In J, the kinetoplast is in early kinetoplast S phase and illustrates the presence of unilateral filaments and attachment to the mitochondrion membrane and kinetoplast. The asterisk in J denotes the "nabelschnur", a filamentous structure rich in basic proteins that links the kDNA discs during their segregation [18]. In G-J scale bar is 250 nm.