Novel Fluorescent Mitochondria-Targeted Probe MitoCLox Reports Lipid Peroxidation in Response to Oxidative Stress In Vivo

Figure 3

Measurements of MitoCLox oxidation in living cells by flow cytometry. MRC5-SV40 cells were incubated with MitoCLox (200 nM) for 1 h before the addition of peroxides. (a) Fluorescence spectra of reduced MitoCLox (blue line) and MitoCLox oxidized by 500 μM H₂O₂ (red line) at 488 nm excitation. The green area is the part of the spectrum detected by the standard bandpass filter 525/40 nm (FL1); the red area is the part of the spectrum detected by filter 575/40 nm (FL2). (b) The ratiometric measurements (FL1/FL2) of MitoCLox oxidation induced by 1 h incubation with H₂O₂. (c) An example of typical histograms obtained in experiments where the cells were treated with H₂O₂ and MitoCLox oxidation was measured as the FL1/FL2 ratio. (d) The time dependence of the MitoCLox oxidation in response to the addition of H₂O₂ (500 μM). (e) Incubation of the cells with mitochondria-targeted antioxidant SkQ1 (20 nM, 2 h) protects the cells from oxidative stress induced by H₂O₂ (500 μM) or 25 μM cumene hydroperoxide (CumOOH). All experiments were triplicated, and each bar represents the ( vs. the control or treated cells without SkQ1).

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