The influence of simultaneous and individual MRB8170/4160 depletion on the steady-state levels of maxicircle-encoded mRNAs. qRT-PCR analysis of pre-edited (P), edited (E), and never-edited (NE) mRNAs encoded on maxicircle DNA. Analysis was performed in the presence or absence of the RNAi-induction agent tetracycline for the double (A) and single knockdowns of MRB8170 (B) and MRB4160 1 (C). For each target amplicon, the relative change in RNA abundance due to the depletion of both or one of the MRB8170/4160 proteins as compared with the untreated controls, which were processed in parallel, was determined by using the cytosolic transcripts β-tubulin and 18S rRNA as internal references. The mean relative abundance of each amplicon is plotted as a bar graph on a logarithmic scale on the y-axis, in which levels above and below the middle axis signify an up- or down-regulation, respectively, while no bar indicates that there was no change in steady-state levels of the transcript upon RNAi silencing; whiskers denote the range of obtained relative abundances within the triplicates. The following pre-edited and edited mRNAs were assayed: ATPase subunit 6 (A6), cytochrome oxidase subunits 2 (co2) and 3 (co3), cytochrome reductase subunit b (cyB), maxicircle unknown reading frame 2 (MURF2), NADH dehydrogenase subunit 7 (ND7), and ribosomal protein S12 (RPS12). The following never-edited RNAs were assayed: cytochrome oxidase subunit (co1) and NADH dehydrogenase subunit 4 (ND4).
