Fig 3.

(A) Cumulative growth of an induced (1 μg/ml Tet) whole-genome RNAi library in bloodstream form T. brucei brucei Lister 427 strain trypanosomes in the presence (squares) or absence (circles) of CpdA. The initial concentration of CpdA was 30 nM, which was increased to 60 nM after 4 days. (B) Relative growth of the surviving RNAi library trypanosome population after selection with CpdA. Cells were grown for 72 h in the presence of 60 nM (+CpdA) or absence of (−CpdA) CpdA with RNAi either induced (+Tet) or uninduced (−Tet). Growth is expressed as a percentage of that of the −Tet, −CpdA population. (C) Ethidium bromide-stained agarose gel (1%, wt/vol) of the genomic PCR products representing the RNAi target fragments in the library constructs selected after 15 days in CpdA (i.e., those fragments that are associated with resistance to CpdA). DNA ladder size markers on the left are denoted in base pairs. Slices refer to the portions of the gel excised for cloning and sequencing.