Fig 1.
Location of images (black box) for immunohistochemical analysis of the cortex, striatum and hippocampus. Modified Nissl sections from The Mouse Brain by Franklin & Paxinos (3rd edition).
Fig 2.
Changes in the ECM are observed in multiple regions of the brain following BCAS.
A) Representative images of collagen IV staining (green) within the cortex, hippocampus and striatum of sham or BCAS-treated (7 and 14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. B) Quantification of collagen IV-positive pixels. N = 4–6 **p < 0.01 C) Representative images of alpha5 staining (red) within the cortex, hippocampus and striatum of sham or BCAS treated (7 and 14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. D) Quantification of alpha5-positive pixels. N = 4–6 *p < 0.05.
Fig 3.
Leakage of Evans blue from blood vessels is observed in multiple regions of the brain following BCAS.
A) Representative images of Evans blue (EB) staining (red) co-labeled with tomato-lectin (green) within the cortex, hippocampus and striatum of sham or BCAS-treated (14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. White arrows indicate example regions of Evans blue leakage from the endothelial cells. B) Quantification of Evans blue-positive pixels. N = 3 *p < 0.05, ***p < 0.001.
Fig 4.
Changes in endothelial tight junctions are observed in multiple regions of the brain following BCAS.
A) Representative images of claudin-5 staining (red) within the cortex, hippocampus and striatum of sham or BCAS-treated (14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. B) Quantification of claudin-5-positive pixels. N = 4–6 C) Quantification of occludin gene expression within the cortex and striatum of sham or BCAS treated (14 day) mice. N = 3 *p < 0.05, **p < 0.01.
Fig 5.
Changes in glial cells are observed in multiple regions of the brain following BCAS.
A) Representative images of GFAP staining (red) within the cortex, hippocampus and striatum of sham or BCAS-treated (14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. B) Quantification of GFAP-positive pixels. N = 4 **p < 0.01, ***p < 0.001 C) Representative images of CD11b staining (green) within the cortex, hippocampus and striatum of sham or BCAS-treated (14 days) mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. D) Quantification of CD11b-positive pixels. N = 4 *p < 0.05.
Fig 6.
Cellular proliferation is increased in multiple regions of the brain following BCAS.
A) Representative images of Ki-67 staining (green) and B) quantification of Ki-67-positive pixels in the cortex, hippocampus and striatum of sham and BCAS-treated (14 days) mice. N = 3 *p < 0.05 Scale bar = 100 um. C) Representative images of Ki-67 staining (green) co-labeled (red) with PECAM (endothelial cells), GFAP (astrocytes), CD11b (microglia) and OSP (oligodendrocytes) within the striatum of 14 day BCAS mice. Scale bar = 100 um. Inset images are a magnified portion to show detail. Scale bar = 300 um. Graph shows quantification of the percentage of Ki-67 positive cells co-labelled with PECAM, GFAP, CD11b or OSP from 14 day BCAS mice. N = 3.
Fig 7.
Decreased number of anastomoses in BCAS-treated mice.
A) Representative cortical images (1x) of Di I perfused brains of sham and BCAS-treated (day 14 and 30) mice. Magnified image below to show detail. White circle is one example of an anastomosis. B) Quantification of the number of anastomoses between the MCA and ACA and the MCA and PCA in sham and BCAS-treated (day 14 and 30) mice. N = 5–8 **p < 0.01 ***p < 0.001 MCA: middle cerebral artery, ACA: anterior cerebral artery, PCA: posterior cerebral artery.