Research Paper Volume 4, Issue 1 pp 28—39

Figure 2. Co-localisation of p53 isoforms with damaged mtDNA. (A) p53^−/− HCT116 cells treated with 100 nM rotenone for 24 hours and stained for 8-oxo-dG and revealed with RPE-conjugated secondary moAb (red fluorescence). Nuclei are counterstained with Hoechst 33258. Punctuated, cytoplasmic red fluorescence indicates that 8-oxo-dG accumulates in mitochondria but not nuclei upon rotenone treatment. (B) flow cytometric detection of 8-oxo-dG after rotenone treatment. C: p53^−/− HCT116 cells transfected with either EGFP-p53R⁷² or EGFP-p53P⁷² pCMS plasmid and treated as in A. Arrows indicate the points in which 8-oxo-dG (red fluorescence) and p53 (green fluorescence) co-localise (yellow dots in the merged picture). Note that in the case of p53R⁷² the green fluorescence is diffused in the cytoplasm and tends to accumulate in mitochondria, while in the case of p53P⁷² the green fluorescence is mainly localised to the nucleus.