FIGURE 6.

Effects of APX3330 treatment on superoxide production and HMGB1 translocation in the myenteric plexus of Winnie mice. A, LMMP preparations of the distal colon were labeled with Mitochondrial superoxide marker MitoSOX (red) indicative of oxidative stress. Increased MitoSOX immunofluorescence was evident in the myenteric ganglia of Winnie sham-treated (n = 5) compared with C57BL/6 control (n = 5) mice (scale bar = 100 µm, 40x magnification). This was alleviated in Winnie APX3330-treated (n = 5) mice (scale bar = 100 µm) (A’) HMGB1 immunoreactivity (green) colocalized with nuclei marker DAPI (blue) in the myenteric plexus. Translocation of nuclear HMGB1 to cytosol (marked with white arrows) was observed in Winnie sham-treated compared with C57BL/6 control mice and was averted in Winnie APX3330-treated mice (n = 5/group; scale bar = 50 µm, 60x magnification). B, MitoSOX fluorescence intensity was assessed relative to ganglion area. C, The number of cells with translocation of HMBG1 from nucleus to cytoplasm was quantified in the myenteric ganglia. Data expressed as mean ± SEM, *P < 0.05, **P < 0.01 compared with C57BL/6 control mice; ^P < 0.05, ^^P < 0.01 compared with Winnie sham-treated mice.