Fig. 3

Characterization of the microglial activation pattern in the postischemic retina upon XBD173 treatment. a Aif1 labeling of retinal flatmounts delineating microglia in the inner plexiform layer representative of cells quantified in the inner retina (INR) and the outer plexiform layer (OPL) at 3 and 14 dps. Scale bars, 20 μm. b Determination of the area occupied by processes of individual microglia as a measure for their activation status. c Microglia were quantified (n = 4 for each condition) in the OPL and INR (e.g., ganglion cell and inner plexiform layer). d Expression levels of Aif1 and Itgam were detected via qPCR. Note that especially 3 days (3) after ischemia, both Aif1 and Itgam expression was the highest in microglia of untreated mice indicative of a stronger microglial activation at this time point Control, c. e Markers for pro-inflammatory M1 microglia were investigated via qPCR in MACS-sorted cells. TNFα (Tnf) expression was similar in both treatment groups except for microglia 14 days (14) post-surgery. In contrast, less F4/80 was expressed in microglia of XBD173-treated mice. f Arginase 1 (Arg1) is a marker for anti-inflammatory, protective M2 microglia. Enhanced expression of Arg1 transcripts may indicate improved neuroprotection in the stressed postischemic retina. b–f Bars represent mean values ± SEM (n = 3–6 mice of each treatment group and time point). */°/•P < 0.05; °°P < 0.01; •••P < 0.001. The color of the circle indicates the treatment group