Figure 16. Vitamin C vs. the DAV triple combination: a mechanistic side-by-side comparison.Left panel: When used as a single agent, Vitamin C can act as a pro-oxidant and induce mitochondrial biogenesis, driving increased mitochondrial protein synthesis and elevated ATP production. Right panel: In contrast, the DAV triple combination would preferentially inhibit the synthesis of proteins that are encoded by mitochondrial DNA (mt-DNA), leading to a strict depletion of essential protein components that are absolutely required for maintaining OXPHOS. In the absence of these required OXPHOS components, this would result in abnormal mitochondrial biogenesis and severe ATP depletion. As predicted, we observed dramatic ATP depletion experimentally. Therefore, Vitamin C amplifies the effects of Doxycycline and Azithromycin, by driving mitochondrial biogenesis, thereby diluting out the pre-existing population of mt-DNA encoded proteins. In summary, this strategy was designed to create a “rho-zero-like” phenotype. Also, since Azithromycin is an established inducer of autophagy, this approach should stimulate mitophagy, to actively eliminate defective mitochondria.