Fig. 2. Hummingbird uncoupling protein (HmUCP) expression in yeast and HmUCP-induced mitochondrial uncoupling. (A) Western blot detection of 12CA5-tagged HmUCP in isolated yeast mitochondria after 12 h of induction with 1 % galactose using anti-12CA5 antibodies. Cells transformed with empty pYES2 vector were used as negative controls. (B) Flow cytometry analysis of mitochondrial membrane potential following the expression of HmUCP in yeast. Rat UCP1 and empty pYES2 vector were used as positive and negative controls, respectively. Yeast cells were incubated in DiOC₆-sensitive dye. The x-axis is a logarithmic scale of fluorescence intensity, and the y-axis represents the number of cells. A shift of the curve towards the left indicates a decreased mitochondrial membrane potential. Reproduced from Vianna et al. (2001) with permission from the American Physiological Society.

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