Fig. 3

Leukocyte adhesion/infiltration, microglial recruitment/activation and gliosis are increased in the retina of P301S mice. a WT and P301S mice were subjected to leukostasis assay at various ages. Bar graph represents the number of leukocytes adherent to the retinal vasculature per retina. n = 5–15/group. b Leukocytes were stained with anti-CD45 antibody in retinal flatmounts of WT and P301S mice, and infiltrated leukocytes in the retina were quantified. n = 5–7/group. c Microglia were stained with anti-Iba1 antibody (purple) at 1 and 3 months of age. Images were taken at the NFL-GCL by confocal microscopy. Squares in the upper panel of images are zoomed in to more clearly show microglial activation. d Bar graphs represent the quantification of morphological parameters of microglia at the NFL-GCL, including soma size and roundness, number and nearest neighbor distance (NND). n = 4–8/group. e Microglia were stained with anti-Iba1 antibody (green) and vasculature was co-labeled with ConA-lectin at 1 month of age. Images were taken at the NFL-GCL by confocal microscopy to show the relationship of microglia and vasculature. Arrows indicate those microglia close to vessels. n = 3/group. f The activation of Müller cells was assessed by immunostaining with antibody against GFAP (red) in retinal sections. Arrowheads indicate activated Müller cells. Blue staining indicates nuclei. n = 3–5/group. Scale bar: 50 µm. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 versus WT. GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer