Figure 3. Localization of TbMyo1 in bloodstream and procyclic forms of T. brucei.

Bloodstream forms were purified from infected blood, fixed and then processed for immunofluorescence using affinity purified anti-TbMyo1 antibodies as described in the methods. The cells were visualized using a Zeiss Axiovert 100 fluorescence microscope. The images were captured and processed using Axiovision software. Bar  = 5 ųm. Panel A. The upper panels present the fluorescence image and reveal the location of TbMyo1 (green) and the nucleus/kinetoplast (blue). The lower panels present the corresponding merge with the phase contrast view. Nonspecific binding was investigated by including the peptide antigen (10 µg.ml⁻¹) along with the α-TbMyo1 affinity purified antibodies. Panel B. A series of images from the same population of bloodstream forms demonstrated that TbMyo1 was present in discrete spots that varied in number, signal intensity and relative location between the nucleus and kinetoplast. Panel C. Analysis of the number of TbMyo I spots in cells. Three different fields of 100 cells were analysed and the results were expressed as mean ± SEM. The analysis revealed a distribution of TbMyo1 spots ranging from 1 to 6 spots per cell. Panel D. Cultured procyclic forms were fixed and processed for immunofluorescence as described in the methods. The upper panels present the fluorescence image and reveal a weak, uniform signal distributed throughout the cell. The lower panels present the fluorescence merged with the corresponding phase image. Nonspecific binding was investigated by including the peptide antigen (10 µg.ml⁻¹) along with the α-TbMyo1 affinity purified antibodies.