Fig. 10

Incubation with MSC-EVs reduces NF-κB activation in hippocampal slices from hyperammonemic rats. A Representative images of immunofluorescence against p50 subunit of NF-κB (green) in hippocampal slices. Nuclei are stained with DAPI (blue). B Ratio of nuclear/cytoplasmic NF-κB p50 subunit in neurons of CA1 region of hippocampus, measured by immunofluorescence and expressed as percentage of control (n = 5–6). C Axial projections of z-stack to confirm p-50 nuclear localization: a representative image showing 3D and 2D projections with its corresponding XZ and YZ planes is shown on the left. Representative images of control and HA samples showing 2D projections with their corresponding XZ and YZ planes are shown on the right. D p65 transcriptional activity in nuclear extracts measured by DNA-binding activity kit. Data of optical density were measured at 450 nm and are expressed as percentage of controls. E Double-immunofluorescence against p50 subunit of NF-kB (green) and Iba1 (staining microglia, in red). Nuclei are stained with DAPI (blue). F Number of microglial cells expressing NF-κB p50 subunit, measured by double immunofluorescence and expressed as cells/mm² (n = 4–6). One-way ANOVA with Tukey post hoc test was performed to compare all groups. Values are the mean ± SEM. Values significantly different from controls are indicated by asterisk (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001), values significantly different from HA group are indicated by a (a = p < 0.05; aaa = p < 0.001; aaaa = p < 0.0001) and values significantly different from HA + EVs group are indicated by b (b = p < 0.05; bb = p < 0.01; bbb = p < 0.001). Sample size of each group is indicated at the bottom of the bars