Fig 1. Screening of RNAi library and identification of TbMCP14.

A) The concentrations of drugs killing 98% (EC₉₈) of T. brucei bloodstream forms were determined using Alamar blue assays. The numbers represent mean values ± standard deviations from 3 independent experiments. B) Time-line of induction of RNAi library, selection of resistant clones, and sequencing of RNAi inserts from resistant parasites. C) Scheme depicting truncated and full-length TbMCP14. The horizontal grey arrows represent two potential TbMCP14 ORFs, encoding proteins of 344 (truncated) and 401 amino acids (full-length). The colored lines at the bottom indicate the alignments of the RNAi inserts isolated from resistant parasites after selection with G25, T3 and T4 (from top to bottom). The right panel shows an ethidium bromide-stained agarose gel with the RT-PCR products amplified using a primer corresponding to the spliced leader sequence (SL) together with the gene-specific primers 1 and 2 indicated below full-length TbMCP14. The numbers on the left indicate the migration of base pair markers. TbMCP14 was the only gene identified in the RNAi screens using either of the three drugs.