Research Article

An Inherited Heteroplasmic Mutation in Mitochondrial Gene COI in a Patient with Prostate Cancer Alters Reactive Oxygen, Reactive Nitrogen and Proliferation

Figure 3

Peroxide and Nitric Oxide are elevated in 6124 mutant cybrid cells. (a) Peroxide levels are elevated in 6124Mut cell lines as measured by flow cytometric analysis of DCF fluorescence. 143B cybrids cell lines containing either the wild type base at position 6124WT or the mutation at position 6124Mut were analyzed. The average DCF fluorescence of five wild type and six mutant clonal cell lines are shown. Mutant cybrids produce significantly more peroxides ( ). Error bars represent the standard error of the mean of 10 data points (WT) and 12 data points (Mut). (b) Mitochondrial superoxide levels are decreased in 6124Mut cell lines compared to 6124WT ( ). The average MitoSOX fluorescence of three wild type and three mutant clonal cell lines are shown. Error bars represent the standard error of the mean of 9 data points each WT and Mut. (c) NO levels are highly elevated in 6124Mut cells compared to 6124WT as measured by DAF-FM Fluorescence ( ). For comparison, 143B cybrids cell lines containing a separate patient mtDNA with either a mutation at position 8993 (8993Mut) or wild type 8993 (8993WT) is shown. Error bars represent the standard error of the mean of 9 data points each 6124WT, 6124Mut, and 3 data points each 8993WT and 8993Mut. (d) Hydroxyl radicals and peroxynitrite anions as measured by hydroxyphenylfluorescein (HPF) remain unchanged. Error bars represent the standard error of the mean of 9 data points each 6124WT and 6124Mut.
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