Figure 6
Effects of the protein levels of DmLRPPRC1 on mitochondrial protein synthesis in Schneider S2 cells. (A) Schneider S2 cells with no plasmid (control) or carrying pMt/Hy (vector) or pMt/DmLRPPRC1/Hy (DmLRPPRC1) were cultured for 5 d in the presence or absence of 0.1 mM CuSO4. Immunoblot analysis was carried out as described in the legend to Fig. 1A. (B) Steady-state levels of mitochondrial transcripts in DmLRPPRC1-overexpressing Schneider S2 cells. Northern blot analysis of the levels of mitochondrial genome encoded transcripts in ClpP-depleted Schneider S2 cells. The nuclear genome encoded transcripts, RP49 and TubA, was used as a loading control. (C) Mitochondrial translation products were labeled in Schneider S2 cells after 5 d of culture in the presence or absence of 0.1 mM CuSO4. Labeling was carried as described in Fig. 1F. Upper panel, aliquots containing 10 μg of total protein each were fractionated by 15–20% gradient SDS-PAGE. Coomassie Brilliant Blue (CBB) staining was used to check for equal loading of the samples. Lower panel, the relative mitochondrial protein synthesis ratio of DmLRPPRC1-overexpressing cells. The relative mitochondrial protein synthesis ratio was quantitated as described in Fig. 1F. The results represent the mean ± SD of three independent experiments and ***P < 0.001 versus control cells without induction (Student’s t-test). Full blots are presented in Supplementary Fig. S9.
