Fig. 6

Analysis of M/M phenotypic markers at different time points after dMCAO. a Coronal sections of the lesioned hemispheres of the inhibitor and control mice brains, at seven (two upper rows of panels) and 14 days (two lower rows) after dMCAO. The sections were triple-immunostained with anti-Iba-1 (red), anti-CD45 (blue), and anti-CD68 (green) antibodies. Representative merged and single-channel images depict part of the infarct core (I, outlined with white dotted lines) and peri-infarct area of stroke. High magnification images demonstrate distribution of Iba-1, CD45, and CD68 in the glial scar area (right panels). Imaging was performed using Zeiss LSM800 Airyscan confocal microscope, using single-scan and tile-scan image acquisitions. Bars: from left to right: 100 and 10 μm. b Quantification of the corrected fluorescence intensity for CD45 and CD68 immunofluorescence staining in the inhibitor (black bars) and control groups (white bar), at 7 and 14 days after dMCAO. N = 5–6 mice per group/per time point, three brain sections per mouse. Error bars: SEM; Student’s t test, *p < 0.05; **p < 0.01. c Representative Western blots and quantification analysis (graphs) of CD206, CD45, CD68, and Iba-1 expression in lysates prepared from the lesioned cortical tissue of the animals from the inhibitor (black bars) and control (white bars) groups, at 7 and 14 days after dMCAO. Graphs: optical density of each band was normalized to β-actin (loading control). N = 5–6 mice per group/per time point. Error bars: SEM; Student’s t test, *p < 0.05