Figure 5. Mitochondrial translation in parasites depleted of guide RNA binding complex (GRBC1/2), RNA editing core complex (MP18/24), RNA editing TUTase 2 (RET2), and knock-in cell lines expressing catalytically active (RET2-CODA WT) or inactive (RET2-CODA D97A) TUTases. RNAi and protein expression were induced for indicated periods of time and translation products were metabolically labeled with EasyTag mix (PerkinElmer Life Sciences) in isotonic buffer supplemented with 0.1 mg/ml of cycloheximide. Cells were collected by centrifugation, dissolved in SDS gel loading buffer, and fractionated by two-dimensional electrophoresis. Gels were stained with Coomassie brilliant blue R250 (inset panels) and exposed to X-ray film (large panels). Based on previous protein identifications,²³ major spots represent COI and Cyb proteins encoded by unedited and edited mRNAs, respectively.