Fig. 4

ASC-EVs treatment modulates the neuroinflammation in SMA mice. (A) The graph shows the quantification of the percentage of astrogliosis (GFAP + signal) in the ventral horns of L1-L5 spinal cord for SMA-PBS (white), SMA-EVs (grey) and WT (black) mice at P10. The ASC-EVs administration significantly decreased the percentage of GFAP-immunopositive profiles in SMA-EVs mice compared to SMA-PBS ones (One-way ANOVA **p = 0.0040; ****p < 0,0001). (B) Representative confocal images showing GFAP+ (red) cells in the ventral horns of PBS- (SMA-PBS) and ASC-EVs treated (SMA-EVs) SMA mice and WT mice. Cell nuclei are labelled by DAPI staining (blue). Scale bar 50 μm. (C) The graph shows the quantification of the percentage of microglial cells (IBA-1 + signal) in the ventral horns of L1-L5 spinal cord for SMA-PBS (white), SMA-EVs (grey) and WT (black) mice at P10. No differences in the percentage of IBA-1 immunopositive profile were observed between SMA-PBS, SMA-EVs and WT mice (One-way ANOVA p > 0.05). (D) Representative confocal images showing IBA-1+ (red) cells in the ventral horns of PBS- (SMA-PBS) and ASC-EVs treated (SMA-EVs) SMA mice and WT mice. Cell nuclei are labelled by DAPI staining (blue). Scale bar 50 μm. (E) The graph displays the microglial cell classification in ramified, bushy or amoeboid, based on their shape; the results are expressed as a percentage on the total number of IBA-1-positive cells for SMA-PBS (white), SMA-EVs (grey) and WT (black) group (Two-way ANOVA *p < 0.05; **p < 0.0050). (F) Representative confocal images showing IBA-1+ (red) microglial cells “ramified”, “bushy” and “amoeboid” in the ventral horns of spinal cord. Cell nuclei are labelled by DAPI staining (blue). Scale bar 50 μm