Figure 3.
DmSUV3 deficiency leads to increased mRNA stability. (A) Southern blot analysis of mtDNA levels in dmsuv3 KD (w;UAS-dmsuv3-RNAi/+;daGAL4/+) and control (w;UAS-dmsuv3-RNAi/+; and w;;daGAL4/+) larvae at 5 days ael. (B) Quantification of mtDNA steady-state levels in control and dmsuv3 KD larvae at 5 days ael. Loading of gels was normalized to nuclear DNA levels detected with a probe against histone. (C) Northern blot analysis of the steady-state levels of mitochondrial mRNAs and rRNAs in KD and control larvae at 5 days ael. (D) Quantification of mitochondrial mRNA and rRNA steady-state levels in control and dmsuv3 KD larvae at 5 days ael. Loading of gels was normalized to the transcript encoding the nuclear ribosomal protein 49. (E) qRT-PCR of mitochondrial mRNAs in KD and control larvae at 5 days ael. RP49 transcript was used as endogenous control. (F) De novo mitochondrial transcription in isolated mitochondria from control and dmsuv3 KD larvae at 5 days ael. Mitochondrial rRNA 12S was used as a loading control. All data are represented as mean ± SEM. (*P < 0.05, **P < 0.01, ***P < 0.001, n = 5).