Figure 1.
Ad.1891 infection results in the selective elimination of mutant mtDNA. (a) The structure of the Ad.1891 expression unit. PCMV(CuO), cumate-inducible cytomegalovirus promoter; NLS, nuclear localization sequence; FLAG, FLAG-tag; M.SmaI, codon-optimized M.SmaI; IRES, internal ribosome entry site; cox, the mitochondrial targeting sequence of the human cytochrome c oxidase subunit VIII; Myc, myc-tag; R.XmaI, codon-optimised R. XmaI. (b) The dose–response of the M.SmaI and R.XmaI expression. MOI, multiplicity of infection; α-FLAG, expression of the FLAG-tagged M.SmaI; α-myc, expression of the myc-tagged R.XmaI; β-actin, loading control; arrow denotes the position of the R.XmaI-specific band. (c) The subcellular localization of R.XmaI. JCP239 cells were plated on coverslips and processed as described in the Materials and methods. α-myc, green staining for the myc-tagged R.XmaI; MT, MitoTracker red; overlay, the superimposition of the previous two images. Yellow, regions of colocalization of R.XmaI and MitoTracker red.