Fig. 3.

Establishment of Cox6b1-3T3 cells. a Quantitative PCR analysis of the mRNA expression levels of Cox subunits, normalized for β-actin mRNA, in control NIH-3T3 cells (Con-3T3) and Cox6b1-overexpressing NIH-3T3 (Cox6b1-3T3) cells. The results are presented as mean ± SEM (n = 6, *p < 0.05 vs. Con-3T3). b Immunoblotting for Cox1, Cox4, and Cox6b1 in the cytosolic and mitochondrial fractions of Con-3T3 and Cox6b1-3T3 cells. β-actin and GAPDH were used as cytosolic markers and VDAC/Porin was used as a mitochondrial marker. c Relative expression of Cox6b1 as determined by densitometry of the immunoblots. Results are presented as mean ± SEM (n = 4, ***p < 0.001 vs. Con-3T3). The values were normalized to those of VDAC/Porin. d Localization of Cox6b1 and MitoTracker Red signals in Cox6b1-3T3 cells. Confocal microscopic images of Cox6b1-3T3 cells stained with MitoTracker Red (mitochondria) and TO-PRO-3 (nuclei). Microscopic images of MitoTracker Red signals, Cox6b1 cells, and the merged signals (TO-PRO-3 and MitoTracker Red) are shown in the left, middle, and right panels, respectively