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Fig. 3. An MPM2-positive structure is retained on the old flagellum post cytokinesis enabling distinction between procylic cells post division. (A) Immunofluorescence image of a representative 1K1N procyclic trypanosome labelled with the anti-phospho-epitope antibody MPM2 (green), showing a focus of MPM2 labelling (FC remnant) at the defined stop point of FC migration. Nuclear and kinetoplast DNA are labelled with DAPI (blue). (B) Quantitative analysis of FC migration as revealed by MPM2 labelling of procyclic cells. The graph plots FC migration (as a ratio of old flagellum length) against new flagellum length in biflagellate cells (blue diamonds). The position of the FC remnant on the old flagellum for a population of 1K1N cells (red squares) is overlaid to demonstrate that the observed focus of MPM2 on 1K1N cells precisely correlates with the defined FC stop point. The reduced overlap between the datasets is a reflection of under representation of very late division cells in the biflagellate population. (C) Cartoon summarising the correlation between the temporal and spatial positioning of the new and old flagellum, the FC and inter basal body separation as deduced from the cell measurement data shown in Fig. 1 and supplementary material Fig. S1. This cartoon also depicts the asymmetric MPM2-staining pattern predicted for 1K1N cells following cytokinesis. DIVN, dividing nucleus; K, kinetoplast; N, nucleus. Bar, 10 µm.
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