1. Lysosomal duplication and segregation defects after TbCALM depletion.
Lysosomal duplication and segregation defects after TbCALM depletion. (A) Duplication and segregation of the endosomal system through the cell cycle in parental cells (control). Incubation in the continuous presence of ConA (top panels, green) labels the entire endosomal system. ConA-labelled structures duplicate and segregate in synchrony with kinetoplast duplication until in pre-cytokinesis (i.e. two kinetoplasts and two fully segregated nuclei) two endosomal systems are visible and apparently both competent for endocytosis. p67-positive lysosome (bottom panels, red) apparently undergoes binary duplication and segregation, concurrent with the endocytic system as a whole. (B) TbCALM depletion causes distension of the p67- (red) and ConA- (green) positive compartment, and retardation or block of segregation (two examples are shown). (C) Thin-section transmission electron microscopy of TbCALM-depleted cells (48 h post RNAi induction). The cytokinesis defect is demonstrated by a hugely distorted cell containing two flagellar pockets (FP) and two nuclei. The inset shows a higher magnification of vacuolar structures (indicated by asterisks) suggested to be lysosomal. When present at the thin-section across a flagellar pocket, CCPs show normal morphological appearance (arrowheads).
Paul T. Manna et al. J Cell Sci 2015;128:
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