Figure 3

Use of real-time quantitative PCR to identify nuclear-coded genes regulating age-associated mitochondrial respiration defects. (a) Comparison of mRNA levels of the candidate six genes in the young and elderly groups. The six gene candidates for regulation of age-associated respiration defects were selected by using gene ontology terms and a microarray heatmap (Supplementary Fig. 4b). Black and open bars are the average gene expression levels of fibroblasts from young and elderly groups, respectively. The young group consisted of fibroblast lines TIG3S (fetus), TIG121 (age 8 months), TIG120 (6 years) and TIG118 (12 years). The elderly group consisted of lines TIG106 (80 years), TIG107 (81 years), TIG101 (86 years) and TIG102 (97 years). Levels of transcripts were normalized against UBC expression. The results of each fibroblast line were shown in the Supplemental Figure 5. Of the six genes examined, age-associate regulation was confirmed to be present in MRPL28, EHHADH and GCAT. (b) Comparison of mRNA levels of MRPL28, EHHADH and GCAT in original and reprogrammed fibroblasts. R3S, R121, R107 and R102 represent fibroblasts reprogrammed from the original fibroblasts TIG3S, TIG121, TIG107 and TIG102, respectively. Levels of transcripts were normalized against UBC expression. Black and open bars are fibroblasts from young and elderly subjects, respectively. Gray bars represent reprogrammed fibroblasts. Experiments were performed in triplicate; error bars indicate ± SD. *P < 0.05.