Research Paper Volume 7, Issue 10 pp 869—881

Figure 2. Mitonuclear OxPHOS imbalance and mitochondrial stress response is elicited in white and beige adipocytes upon starvation. (A) OxPHOS protein ratio (right panel) evaluated by calculating the ratio between ^mtDNA-and ⁿDNA-encoded OxPHOS proteins after Western blot (left panel) followed by densitometric analysis. (B) OxPHOS gene expression ratio evaluated by RT-qPCR. (C) OxPHOS protein ratio evaluated as described in (A). (D) OxPHOS gene expression ratio evaluated by calculating the ratio between ^mtDNA-(MTCO1 and ATP6) and ⁿDNA-encoded (SDHA and Cox4b) OxPHOS mRNAs. (E, F) Mitochondrial stress response assessed by Western blot analysis of SOD2 (E) and through RT-qPCR of ClpP (F). (G) OxPHOS gene expression ratio evaluated as described in (D). (H) mRNA levels of ClpP measured by RT-qPCR. (I, J) OxPHOS protein ratio (I, left panel) evaluated as described in (A). SOD2 protein (I) and mRNA levels of ClpP (J) evaluated by Western blot and RT-qPCR, respectively (n=4 mice per group). (K) OxPHOS gene expression ratio evaluated as described in (D) (n=4 mice per group). Actin and TOMM20 served as loading controls. Bar graphs are expressed as mean ±S.D. (n=4; *p<0.05; **p<0.01 vs CM or ad libitum fed mice; °p<0.05). NR: nutrient restriction; CM: complete medium.