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

Figure 7

Analysis of MitoCLox oxidation in the culture of myoblast MB135. (a) Myoblasts were seeded in 35 mm dishes at an initial density of cells/well and cultured for 4 days. Tempol (0.1 mM) was added at the beginning of culturing. Then, 200 nM MitoCLox was added for 5 h, and cells were analyzed using a Nikon Eclipse Ti (Nikon) confocal microscope with excitation at 488 and at 562 nm. Higher magnification was used to reveal mitochondrial morphology. (b) Myoblasts were seeded and cultured as in (a). Then, MitoCLox (200 nM) was added for 5 h, and cells were analyzed using the FC 500 flow cytometer in green (FL1) and red (FL2) channels. (c) Myoblasts were seeded in 6-well cell culture plates (9.6 cm² surface area per well) at initial density cells/well and cultured for 4 days. Then, 200 nM MitoCLox was added for 5 h, and cells were analyzed as in (b). The cell fraction with oxidized MitoCLox was measured. Phase-contrast images of the final cultures seeded at different densities are shown.

(a)

(b)

(c)