Figure 1. Rab11 is essential for proliferation of bloodstream T. brucei cells.
The TbRab11 RNAi cell line was grown in the presence of tetracycline (1 μg/ml) to induce specific knockdown. A. Cell density was measured and plotted against time (mean ± std. dev., n=3 biological replicates). Cultures were diluted to starting density every 24 hours. Empty circles, tet− control cells; closed circles, tet+ silenced cells. B. The extent of TbRab11 mRNA knockdown was assayed by real-time qRT-PCR at the indicated time points. Values are normalized to uninduced controls (mean ± std. dev., n=3 biological replicates). All subsequent experiments were performed at 22 hr of TbRab11 silencing. C. Immunofluorescence microscopy was performed with fixed permeabilized cells using anti-BiP (green) and anti-p67 (red) to localize the ER and lysosomes respectively. Nuclei and kinetoplasts were stained with DAPI (blue). Representative 3-channel images are presented. Bar indicates 2 μm.