Rats were sacrificed 35 days after MCAO. Infarct volume was measured on 7 hematoxylin and eosin (H&E) stained coronal brain sections as previously described12 (link). DM rats without MCAO were sacrificed 65 days after NTM-STZ injection, whereas age-matched Non-DM control rats were sacrificed after the final behavioral tests.
Immunofluorescent staining was performed on brain coronal sections, according to our published protocols15 (link). The following primary antibodies were used: mouse anti-BrdU (Dako) for proliferating cells, goat anti-doublecortin (DCX, Santa Cruz) for neuroblasts, rabbit anti-NG2 (Millipore) for oligodendrocyte progenitor cells (OPCs), mouse anti-adenomatous polyposis coli (APC, Abcam) for mature oligodendrocytes, mouse anti-phosphorylated neurofilament heavy chain (pNFH, Affinity Bioreagents) for axons and dendrites, rabbit anti-myelin basic protein (MBP, Millipore) for myelin, rabbit anti-glial fibrillary acidic protein (GFAP, Dako) for astrocytes, rabbit anti-aquaporin-4 (AQP4, Abcam), mouse anti-endothelial barrier antigen (EBA, Sternberger Monoclonals), and goat anti-fibrin/fibrinogen (Accurate Chemical& Scientific). The Golgi-Cox impregnation method along with the FD Rapid Golgi Stain kit (FD NeuroTechnologies) was used to identify Golgi-Cox impregnated dendrites and spines16 (link). Please see Supplemental Data for details.