Research Paper Volume 5, Issue 11 pp 835—849

Figure 7. Sirt4 regulates expression of nuclear encoded mitochondrial genes in a feedback loop via ANT2/AMPK/PGC1α signaling. (A–C) Quantitative RT-PCR of nuclear encoded mitochondrial genes (A) TFAM, (B) NRF1 and (C) Cytochrome C from RNA isolated from HEK293T cells transfected with control, Sirt4-FLAG, ANT2-siRNA and Sirt4-FLAG/ANT2-siRNA constructs. (D) Western blots for TFAM, Porin, Cytochrome C, Sirt4-FLAG and actin in control and Sirt4OE HEK293T cells. (E) Quantitative RT-PCR of mitochondrial DNA (normalized to nuclear DNA) in DNA isolated from HEK293T cells transfected with control, Sirt4-FLAG, ANT2-siRNA and Sirt4-FLAG/ANT2-siRNA constructs. Statistical significance was calculated using ANOVA: * p < 0.05, ** p < 0.01, *** p < 0.001 or as indicated. Error bars indicate mean values ± SEM.