Fig 2.

Inducible RNAi-mediated ablation of TbPPR9. (A) Representative growth curve of tetracycline (Tet)-uninduced (−Tet) and -induced (+Tet) representative clonal T. brucei TbPPR9 RNAi cell line in standard culture medium SDM-79. (B) Northern analysis of total RNA isolated from the TbPPR9 RNAi cell line induced for the indicated times. The Northern blot on the top was probed with a labeled DNA fragment covering the region of the mRNA that is targeted by the RNAi (RNAi insert), whereas the blot on the bottom was probed with a labeled DNA fragment that recognizes a region in the TbPPR9 mRNA outside the region that is targeted by the RNAi (outside RNAi region). In both cases, ethidium bromide stains of the rRNA region are shown as loading controls. (C) Combination of tetracycline-inducible expression of the Ty1 tagged TbPPR9 and the tetracycline-inducible TbPPR9 RNAi in the same cell line. Total proteins were analyzed by immunoblotting at the indicated time points after tetracycline addition using the BB2 antiserum or KDH antiserum (KDH serves as a loading control).