Immunohistochemical staining of DPIV, DP8 and APN in ipsilateral cortices at different time points after eMCAO. Confocal fluorescence images were captured either 3 (A-C) or 7 days (D-I) post eMCAO. DAPI was used to stain cell nuclei (A-I, blue fluorescence). At day 3, the time point when microglia activation peaked, DPIV (A, green florescence) was co-stained with IB4 (red) to indentify immunoreactive microglia, whereas DP8 (B, green) and APN (C, red) were co-labeled with ED1 (B, red; C, green), a marker of reactive macrophages. Co-localization of DPIV, DP8 or APN labeling with IB4 (activated microglia) and ED1 (active microglia/macrophages), respectively, is visualized by merged fluorescence signals (A-C, yellow, arrows). At day 7, DPIV (D, G, red), DP8 (E, H, green) and APN (F, I, green) were co-stained with Neurotrace targeting neuronal prerikarya (D, green) or NeuN, labeling neuronal nuclei (E, F, red). Alternatively, DPIV, DP8 and APN were co-labeled with GFAP, expressed in astroglial cells (G, green; H and I, red). Co-localization of each protease with the respective cell markers is seen (D-I, yellow, arrows). Scale bars = 10 μm (A-F) and 20 μm (G-I).