Goat anti cd31

Brain and tumor tissues were collected and placed in 10% formalin under constant rocking at 4 °C for ~48 h. Next, tissue was transferred to 30% sucrose (Sigma-Aldrich), 0.01% sodium azide (Sigma-Aldrich) and stored in the cold room under constant rocking for at least 24 h. Tissue was cryo-sectioned at 30 μm thickness. For DNA damage staining, tissue slides were incubated with rabbit or mouse anti-phospho-ser139 histone γH2AX antibody (Cell Signaling Technologies) at a dilution of 1:100. Cleaved-caspase 3 staining was performed in tissue slides with incubation of a rabbit anti-cleaved-caspase 3 (Cell Signaling Technologies) antibody at a dilution of 1:100. For astrocyte staining, tissue slides were incubated with mouse anti-GFAP (Cell Signaling Technologies) at a dilution of 1:300. Endothelial cells were stained with goat anti-CD31 (Abcam) at a dilution of 1:100. Neurons were stained with rabbit NeuN (Cell Signaling Technologies) at a dilution of 1:300. Next, slides were incubated appropriately with either goat anti-rabbit or anti-mouse antibody conjugated with Alexa Fluor 647 (Jackson Laboratory) at a dilution of 1:500, donkey anti-mouse, anti-rabbit or anti-goat antibody conjugated with Alexa Fluor 488 (Jackson Laboratory) and 1:1000 of Hoechst 33342 (Thermo Fisher Scientific) for nuclear staining. Slides were visualized via confocal microscopy.