Fig. 3

Blocking of platelet GPIb diminished ultra-early T-cell recruitment in the ischemic brain. a Representative immunocytologic stainings (top) and quantification (bottom) of brain-infiltrating CD4-positive T lymphocytes (Cy3, red), nuclei (DAPI, blue) in the whole ipsilateral hemisphere 2, 3, and 4 h after MCAO or after 2 h of MCAO with additional 6 h of reperfusion in mice treated with rat IgG Fab (Ctrl Fab) or p0p/B Fab (a-GPIb Fab) using × 20 objective. Scale bar 100 μm (n = 4–5). b (top) Native whole mouse brain with schematic overview of ACA and MCA territories. The ROI used for T-cell quantification is shown on a TTC-stained coronal brain section (2 h MCAO). (bottom) Quantification of brain-infiltrating CD4-positive T lymphocytes (Cy3, red), nuclei (DAPI, blue) in the ROI at 2, 3, and 4 h after MCAO in mice treated with rat IgG Fab (Ctrl Fab) or p0p/B Fab (a-GPIb Fab) using × 20 objective (n = 5). Representative images (left) and quantification (right) of c CD8a-positive cells (CD8a (Alexa 647, red), nuclei (DAPI, blue), d Ly-6B.2-positive cells, or e CD11b-positive cells (3,3-Diaminobenzidin (DAB)) in the ipsilateral hemisphere 2, 3, and 4 h after MCAO or after 2 h of MCAO with additional 6 h of reperfusion in mice treated with rat IgG Fab (Ctrl Fab) or p0p/B Fab (a-GPIb Fab) using × 20 objective. Scale bar 100 μm (n = 4–5). *P< 0.05, **P< 0.01 between the indicated groups, 2-tailed Student t test or, in the case of nonparametric functional outcome, the Wilcoxon–Mann–Whitney U-test was applied. ACA, anterior cerebral artery; MCA, middle cerebral artery; ROI, region of interest