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Figure 4


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TbKINX1B is involved in basal body/FPC segregation in PCF. A. Left panel, Western blot analysis of whole cell protein extracts from PCF TbKINX1BRNAi cell line in the presence (+) or absence (−) of tetracycline after 24 h, 48 h or 72 h probed with anti-TbKINX1B, anti-TbBILBO1, and anti-Tubulin (TAT1) as a loading control (left panel). Right panel, PCF growth curves showing that induction of RNAi down-regulation of TbKINX1B (TbKINX1BRNAi) does not dramatically affect cell growth when compared to WT and non-induced cells (n = 3). B. Down-regulation of TbKINX1B induces misplaced kinetoplasts. Left panel, IFA micrographs from CSK preparation of WT (a) and TbKINX1BRNAi cell lines after 48 h of induction (b, c). Kinetoplasts (K) and nuclei (N) were stained with DAPI. Right panel, quantification of kinetoplast localization in 1K1N WT cells and in TbKINX1BRNAi cells after 24 h, 48 h and 72 h of induction. C. Left panel, immunolocalization of TbKINX1B (green) in TbBILBO1RNAi induced detergent-extracted cells. Right panel, immunolocalization of TbBILBO1 (green) in TbKINX1BRNAi induced detergent-extracted cells. Scale bars 5 μm.

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