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Figure 1.

βL activates AMPK signalling pathway.

(A) βL increases NAD+/NADH ratio by acute injection. NAD+/NADH ratio was calculated in the tissue of mice intravenously with vehicle (n = 5) or 5 mg/kg βL (n = 5) (B) Effect of βL on phosphorylation of AMPK for EDL (n = 5 per each group). Values represent mean± SEM.

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Figure 1 Expand

Figure 2.

βL decreases body mass.

(A) Body weight was measured over 10 months for all survivors. (B) Food intake was measured and calculated as average daily values for all survivors. (C) Fat mass was measured after 10months treatment (n = 5) (D) Changes in body temperature after injection with vehicle (n = 4) or 5 mg/kg βL (n = 4). Values represent mean±SEM. *p<0.05 **p<0.001 versus control #p<0.05 ##p<0.001 versus CR.

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Figure 2 Expand

Table 1.

Effects on metabolic parameters in plasma.

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Table 1 Expand

Figure 3.βL.

βL increases energy expenditure.

(A) Energy expenditure as measured by changes in VO2 consumption in indirect calorimetry over 12 hr (n = 7 for each group). (B) Respiratory quotient (RQ) was calculated from the ratio of VCO2/VO2. Data represent the mean ± SEM. *p<0.05 **p<0.001 versus control #p<0.05 ##p<0.001 versus CR.

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Figure 3.βL Expand

Figure 4.

βL delays functional decline.

(A)Total distance (B) Measurement of freezing time for cued conditioning (C)Time to fall from an accelerating rotarod (D)Peak tension measurements of grip strength (E)Number of successful trials in pole tests (n = 10–15 for each group) for control (C), calorie restriction (CR), and β-lapachone supplemented (βL) mice. Data represent the mean ± SEM. *P<0.05 versus control; **P<0.001 versus pre-CS.

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Figure 5.

βL preserves mitochondria.

(A) Electron microscopy (×15,000 magnification) of skeletal muscle. Scale bar represents 2 µm. (B) Quantification of inter-myofibrillar mitochondria number per 100 µm2 in the skeletal muscles after 10 months treatment (analysis of seven images per mouse, n = 5 per group) (C) Quantitative RT-PCR of genes related to mitochondrial function (n = 4 mice/group). Data represent mRNA levels relative to cyclophilin A. Data represent the mean ± SEM. *P<0.05 versus control.

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Figure 6.

βL shifts expression patterns towards those for calorie restriction.

(A) Parametric analysis of gene-set enrichment (PAGE) (up-regulated (red) or down-regulated (blue)) (B)(upper) Venn diagram shows number of overlapping genes between data sets.(lower) Gene ontology analysis. Results are plotted as negative log-transformed p- values.

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Figure 6 Expand

Table 2.

The most highly up-regulated and down-regulated genes in skeletal muscle.

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Table 2 Expand