QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. TbLC1 localizes along the flagellum and is required for motility. (A) Fluorescence microscopy of live cells and cytoskeletons from TbLC1-GFP strains shows that TbLC1-GFP is localized to the flagellum in induced cells (+Tet). There is background autofluorescence in the cell body of induced and uninduced (–Tet) live cell samples that is variable (Baron et al., 2007). Cytoskeletons were stained with DAPI (blue) to visualize the nucleus and kinetoplast relative to TbLC1-GFP (green). (B) Western blot analysis with anti-GFP antibody of TbLC1-GFP cellular fractions from induced (+Tet, 24 hpi) and uninduced (–Tet) cells confirms that TbLC1 is stably associated with the flagellum. L, lysates; S1, detergent-soluble proteins; P1, cell cytoskeletons; S2, NaCl soluble proteins; P2, flagellar cytoskeletons. The same fractions were blotted with anti-trypanin (TPN) monoclonal antibody as a loading control (bottom panel). (C) Northern blots of RNA from an uninduced (–Tet) and induced (+Tet, 24 hpi) TbLC1 RNAi knockdown strain probed with TbLC1 and TbCMF46 (control) DNA fragments. (D) Images of whole cultures of uninduced (–Tet) and induced (+Tet) TbLC1 knockdown cells over a 5-day induction. At 48 hpi multicellular clusters appear in the culture containing 5-10 cells each, and increase in size and number over time. (E) Growth curve of uninduced (–Tet) and induced (+Tet) TbLC1 cells over a 5-day period. (F) Sedimentation curves (Baron et al., 2007; Bastin et al., 1999; Ralston et al., 2006) for the uninduced (–TET) and induced (+TET, 24 hpi) cells. Error bars show the s.d. for three experiments.

Return to article