Journal: Physiological Reports

Article Title: Chronic consequences of ischemic stroke: Profiling brain injury and inflammation in a mouse model with reperfusion

doi: 10.14814/phy2.16118

Figure Lengend Snippet: Study design and experiment timeline. Following a 5–7‐day acclimation period to their housing environment, mice underwent functional activity assessment (a), followed by a surgical procedure to induce transient middle cerebral artery occlusion (b). Following ischemic stroke, functional activity assessment, and magnetic resonance imaging (MRI) were carried out at 2 and 30 days poststroke (c and d) after which plasma and tissue (e) were collected at 30 days postinjury. Plasma was assessed for NFL concentration (f) and brain tissue (g) was processed for immunohistochemistry to detect microglia (IBA1) astrocytes (GFAP), B lymphocytes (B220), and T cells (CD3ε). BODIPY was used to assess lipid accumulation. Created with BioRender.com (YZ26KHM453).

Article Snippet: Primary antibodies against CD3ε (1:1000; BD Biosciences, Cat. No. 550277; RRID:AB_393573) and B220/CD45R (1:500; BD Biosciences, Cat. No. 553085; RRID:AB_394615) were used with the appropriate secondary antibody and visualized using the VECTASTAIN Elite ABC Reagent, Peroxidase, R.T.U. (Vector Laboratories, Cat. No. PK‐7100) and Vector DAB Substrate (3,3′‐diaminobenzidine) Kit (Vector Laboratories, Cat. No. SK‐4100).

Techniques: Functional Assay, Activity Assay, Magnetic Resonance Imaging, Concentration Assay, Immunohistochemistry

Journal: Physiological Reports

Article Title: Chronic consequences of ischemic stroke: Profiling brain injury and inflammation in a mouse model with reperfusion

doi: 10.14814/phy2.16118

Figure Lengend Snippet: T cells populate the infarct in male and female mice at 30 days poststroke (DPS). Stitched images of CD3ε (T cells) immunostaining in tissue from male (a) and female (b) mice at 30 DPI and summary data of the measured percent area of CD3ε immunoreactivity within the ipsilateral hemisphere (male n = 4; female n = 6). Scale bar: 500 μm.

Article Snippet: Primary antibodies against CD3ε (1:1000; BD Biosciences, Cat. No. 550277; RRID:AB_393573) and B220/CD45R (1:500; BD Biosciences, Cat. No. 553085; RRID:AB_394615) were used with the appropriate secondary antibody and visualized using the VECTASTAIN Elite ABC Reagent, Peroxidase, R.T.U. (Vector Laboratories, Cat. No. PK‐7100) and Vector DAB Substrate (3,3′‐diaminobenzidine) Kit (Vector Laboratories, Cat. No. SK‐4100).

Techniques: Immunostaining

Journal: Physiological Reports

Article Title: Chronic consequences of ischemic stroke: Profiling brain injury and inflammation in a mouse model with reperfusion

doi: 10.14814/phy2.16118

Figure Lengend Snippet: Study design and experiment timeline. Following a 5–7‐day acclimation period to their housing environment, mice underwent functional activity assessment (a), followed by a surgical procedure to induce transient middle cerebral artery occlusion (b). Following ischemic stroke, functional activity assessment, and magnetic resonance imaging (MRI) were carried out at 2 and 30 days poststroke (c and d) after which plasma and tissue (e) were collected at 30 days postinjury. Plasma was assessed for NFL concentration (f) and brain tissue (g) was processed for immunohistochemistry to detect microglia (IBA1) astrocytes (GFAP), B lymphocytes (B220), and T cells (CD3ε). BODIPY was used to assess lipid accumulation. Created with BioRender.com (YZ26KHM453).

Article Snippet: Primary antibodies against CD3ε (1:1000; BD Biosciences, Cat. No. 550277; RRID:AB_393573) and B220/CD45R (1:500; BD Biosciences, Cat. No. 553085; RRID:AB_394615) were used with the appropriate secondary antibody and visualized using the VECTASTAIN Elite ABC Reagent, Peroxidase, R.T.U. (Vector Laboratories, Cat. No. PK‐7100) and Vector DAB Substrate (3,3′‐diaminobenzidine) Kit (Vector Laboratories, Cat. No. SK‐4100).

Techniques: Functional Assay, Activity Assay, Magnetic Resonance Imaging, Concentration Assay, Immunohistochemistry

Journal: Physiological Reports

Article Title: Chronic consequences of ischemic stroke: Profiling brain injury and inflammation in a mouse model with reperfusion

doi: 10.14814/phy2.16118

Figure Lengend Snippet: B lymphocytes populate the infarct in male and female mice at 30 days poststroke (DPS). (a) Stitched images of B220 (B lymphocytes) immunostaining in tissue from male (a) and female (b) mice along with summary data of the measured percent area of B220 immunoreactivity within the ipsilateral hemisphere at 30 DPI (male n = 4; female n = 6). Scale bar: 500 μm.

Article Snippet: Primary antibodies against CD3ε (1:1000; BD Biosciences, Cat. No. 550277; RRID:AB_393573) and B220/CD45R (1:500; BD Biosciences, Cat. No. 553085; RRID:AB_394615) were used with the appropriate secondary antibody and visualized using the VECTASTAIN Elite ABC Reagent, Peroxidase, R.T.U. (Vector Laboratories, Cat. No. PK‐7100) and Vector DAB Substrate (3,3′‐diaminobenzidine) Kit (Vector Laboratories, Cat. No. SK‐4100).

Techniques: Immunostaining