FIGURE 4.

Analysis of intracellular localization of ROS by a redox sensor PF-H₂TMRos. A and B, HT22 cells were treated with 5 mm glutamate for an indicated time. The cells were incubated with 5 μm PF-H₂TMRos and 5 nm LysoSensor (A) or with 5 μm PF-H₂TMRos and 50 nm MitoTracker (B). Generated fluorescent signals were visualized by confocal microscopy with constant fluorescent parameters. Merged images are shown on the right. Experiments were repeated four times with reproducible results. C, HT22 cells transfected with damage-regulated autophagy modulator (DRAM)-EGFP or M6PR-EGFP were incubated with 5 μm PF-H₂TMRos for 30 min. Intrinsic EGFP signals and PF-TMRos fluorescent signals were simultaneously observed with confocal microscopy. Merged images are shown on the right. Bar, 10 μm.