Zinc Protects Oxidative Stress-Induced RPE Death by Reducing Mitochondrial Damage and Preventing Lysosome Rupture

Figure 2

CellROX staining of oxidant generation in RPE following oxidative stress and zinc depletion. RPE cells under different treatment conditions were incubated with 5 μM per well concentration of CellROX for 30 min at 37°C, followed by MitoTracker and Hoechst 33,342. Cells were washed and fresh medium was added to the wells and imaged by confocal microscopy (a–f) or analyzed by a fluorescent plate reader (g). (a) Control RPE cells in 10% serum. (b) oxPOS-treated RPE cells in 10% serum. (c) Serum-free RPE cells in 1.5 μM zinc. (d) Serum-free RPE cells in 1.5 μM zinc + oxPOS. (e) 15 μM zinc-supplemented RPE cells. (f) oxPOS + 15 μM zinc-supplemented RPE cells. Red: MitoTracker; blue: nuclei Hoechst 33,342; and green: oxidant DNA in CellROX. (g) Histogram showing fluorescence intensity of CellROX green in different treatment groups of cells analyzed by a fluorescent plate reader. Mean ± SEM is plotted for 6 replicates for each condition. , , and . Data analyzed by one-way ANOVA followed by Tukey’s multiple comparison test.

(a)

(b)

(c)

(d)

(e)

(f)

(g)