Identification of a Bacterial-Like HslVU Protease in the Mitochondria of Trypanosoma brucei and Its Role in Mitochondrial DNA Replication
Figure 2
RNAi knockdown of TbHslVU affects cell growth and kinetoplast morphology.
(A). Cells were grown in the presence (+) of tetracycline to induce RNAi for 7 days, and cell growth was monitored daily. Northern blots were performed to assess levels of TbHslV, TbHslU1 and TbHslU2 mRNA before (−) and after (+) 2 days of RNAi (insets). (B–E). Un-induced control cells (B) and cells after RNAi induction for 7 days (C) were labeled with YL1/2 antibody for basal bodies (BB, arrowheads) and counterstained with DAPI for nucleus (N) and kinetoplast (arrows). (D). Two TbHslVU RNAi cells at the final stage of cell division were still connected by a thin thread of kinetoplast DNA (arrows) between two basal bodies (arrowheads) in two well-separated cells. Bars: 2 µm. (E). Tabulation of RNAi cells with kinetoplasts in varying sizes and morphologies. Approximately 200 cells were counted at each time point and the data represent averages from three independent experiments.