Figure 10.
DFO blocked gp120-, morphine-, and iron supplementation-induced decreases in mitochondrial levels of H2S. (A, C) Representative confocal images of SH-SY5Y cells pretreated for 1 h with DFO (50 μM) or water (control) and then treated for 1 h with gp120 (500 pM), morphine (1 μM), gp120 plus morphine, and FAC (50 μM) prior to staining for H2S with P3 probe (green), for mitochondria with MitoTracker Red (red), and for nuclei with Hoechst 33342 (blue). Scale bars = 10 μm. Boxed inserts are magnified mitochondria containing H2S. Scale bars = 1 μm. (B, D) Semi-quantitative changes in mitochondrial H2S were determined using Imaris 3D software and data were represented as fold changes in mean fluorescence intensity (MFI) inside MitoTracker-positive mitochondria. Data were shown as (mean ± SD) and individual data points (n = 6–7) were illustrated. Each data point (n) represents MFI of P3 staining inside mitochondria from multiple cells. A minimum of 61 cells per condition from two biological replicates were analyzed. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analyses. ns = non-significant, *p < 0.05, **p < 0.01, ****p < 0.0001.