Figure 2.

(A) Northern blots of RNA from induced and uninduced TbCen1 culture. Membrane containing 12 μg total RNA from 3-d uninduced (Un) and induced (I) cells was used. Membrane for TbCen1, in addition to hybridization with either specific centrin DNA or α-tubulin (Ngo et al., 1998) probes was reprobed with probes of TbCen2-5 to confirm on the specific inhibition of only the TbCen1 transcript. (B) Western blots on the parasite lysates showing reduction in the protein level of TbCen1 during RNAi induction by anti-LdCen1 Ab (top) and the loading control showing the level of α-tubulin by anti-tubulin Ab (bottom). Un, uninduced; In, induced. In each lane 20 μg of total extracted proteins were loaded. (C) The effect of TbCen1 knockdown on the in vitro growth of T. brucei procyclics. The cells were grown with (I) or without (Un) tetracycline. The data represent the means of ± SD of three independent experiments. *p < 0.04. (D) Geimsa-stained images of the control and TbCen1-depleted cells, 4 d after RNAi induction. Scale bar for both images, 5 μm. AF, attached flagella; DF, detached flagella.