Quantitative Proteomics Uncovers Novel Factors Involved in Developmental Differentiation of Trypanosoma brucei
Fig 5
Differentiation defect of ∆DOT1B trypanosomes.
(A) Cell cycle profiles of propidium iodide stained differentiating trypanosomes (flow cytometry). ∆DOT1B cells re-enter the cell cycle normally (t = 13h). Later in differentiation (t = 22h; t = 32h) the profiles show peak broadening indicating the appearance of cells with abnormal DNA content (red arrows) and enucleated “zoids” (asterisks). (B) Indirect immunofluorescence analysis of differentiating ∆DOT1B trypanosomes with karyokinesis defects (t = 22h). Wild-type cells shows normally shaped nuclei and kinetoplasts (a: 1N1K, b: 2N2K). During differentiation ∆DOT1B cells are unable to divide their nucleus properly resulting in cells with two kinetoplasts and one elongated or asymmetrically divided nucleus (c-f). Cytokinesis is not impaired demonstrated by the appearance of cells with asymmetrically divided nuclei and one kinetoplast (g) or anucleated “zoids” (h) (green: tubulin; red: histone 3; blue: DAPI; bars: 1 μm).