Fig. 3 | Nature Communications

Fig. 3

From: Pentatricopeptide repeat poly(A) binding protein KPAF4 stabilizes mitochondrial mRNAs in Trypanosoma brucei

Fig. 3

KPAF4 repression effects on cell growth and polyadenylation complex. a Northern blotting analysis of KPAF4 mRNA downregulation by inducible RNAi. b Growth kinetics of procyclic parasite cultures after mock treatment and KPAF4 RNAi induction with tetracycline. Data representative of three independent experiments are shown as mean ± s.d. c Quantitative real-time RT-PCR analysis of RNAi-targeted KPAF4 mRNA, and mitochondrial rRNAs and mRNAs. The assay distinguishes edited and corresponding pre-edited transcripts, and unedited mRNAs. RNA levels were normalized to β-tubulin mRNA. RNAi was induced for 55 h. Error bars represent the standard deviation from at least three biological replicates. The thick line at “1” reflects no change in relative abundance; bars above or below represent an increase or decrease, respectively. P, pre-edited mRNA; E, edited mRNA. d Cell lysates prepared at indicated time points of KPAF4 RNAi induction were sequentially probed and re-probed by quantitative immunoblotting on the same membrane in the following order: antigen-purified rabbit polyclonal antibodies against KPAP1, KPAF1, KPAF3, GRBC1/2, and mouse monoclonal antibodies against RET1 TUTase. Signals were normalized against three independent loading standards and mean values calculated

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