FIGURE 5.

Determination of the association of SIRT1 and PGC-1α with mtDNA and TFAM by confocal microscopy in HeLa cells. A, mtDNA was labeled by BrdUrd incorporation followed by methanol-acetone fixing and immunostaining with mouse anti-BrdUrd. SIRT1 or PGC-1α was also immunostained using Santa Cruz Biotechnology antibodies. Cells were then visualized by confocal microscopy, and the level of superimposition of SIRT1 or PGC-1α-derived signals with that of BrdUrd was evaluated by calculation of the Pearson's correlation coefficient (R(r)). Values >0.50 were considered to be significant. B, HeLa cells were fixed with paraformaldehyde and immunostained with goat anti-TFAM and rabbit Santa Cruz Biotechnology anti-PGC-1α or SIRT1. Cells were then visualized by confocal microscopy, and the level of superimposition of SIRT1 or PGC-1α-derived signals with that of TFAM was evaluated by calculation of the Pearson's correlation coefficient (R(r)). Values >0.50 were considered to be significant. Scale bar, 10 μm.