Fig. 6

M8I treatment reduces oxidative stress in astrocytes in ouabain-injected mice. A The expression of nitrotyrosine in astrocytes was assessed using immunofluorescence. Brain sections were collected from three groups of mice and subjected to immunofluorescence staining with GFAP (astrocyte marker, green) and nitrotyrosine (oxidative stress marker, red). B–D Tracking (arrows shown in A) of the intensities for nitrotyrosine (red) and GFAP (green) in the brain sections indicated the intensity of nitrotyrosine produced in astrocytes. The analysis of nitrotyrosine intensity revealed an increase in the ouabain-injected mice groups compared to the control or M8I-treated groups. E The quantitative analysis of nitrotyrosine intensity has revealed an increase in the CA1 region of ouabain-injected mice, while M8I treatment reduced the nitrotyrosine intensity in the CA1 region of ouabain-injected mice. Cell intensities were quantified by the QuPath software Cell Detection function. Quantitative data is presented as means ± SEM; and analyzed using one-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001. The cell number counter utilizes Qupath, and the cell counts are as follows: control (182 cells), ouabain (214 cells), and ouabain + M8I (162 cells)