Journal: PLoS ONE

Article Title: Astrocytic Toll-Like Receptor 3 Is Associated with Ischemic Preconditioning- Induced Protection against Brain Ischemia in Rodents

doi: 10.1371/journal.pone.0099526

Figure Lengend Snippet: (A and B) Western blotting showed that protein expression of Toll-like receptor 3 (TLR3) and glial fibrillary acidic protein (GFAP) was elevated at 12 h and 24 h after IPC compared with that in sham-operated controls. (C and F) TLR3 immunoreactivity. (D and G) GFAP immunoreactivity. (E and H) Merged images show that co-localization of TLR3 and GFAP was increased at 24 h after IPC. Values are expressed as mean ± SD; n = 3 per group; * p <0.05. Magnification in C-H: 200x.

Article Snippet: For double staining of TLR3 and glial fibrillary acidic protein (GFAP), the sections were incubated together with goat anti-TLR3 antibody (1∶50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and mouse anti-GFAP antibody (1∶200; Epitomics, Cambridge, UK) overnight at 4–8°C, and then incubated sequentially with fluorescein-labeled secondary antibody for 2 h at room temperature.

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: Astrocytic Toll-Like Receptor 3 Is Associated with Ischemic Preconditioning- Induced Protection against Brain Ischemia in Rodents

doi: 10.1371/journal.pone.0099526

Figure Lengend Snippet: (A and B) Protein expression of TLR3 was upregulated after IPC and OGD, as measured by Western blot analysis (A) and immunofluorescence staining (B). (C and D) IPC of astrocytes before OGD enhanced protein expression of TRIF (C) and pIRF3 (D). (E) IPC prevented the OGD-induced upregulation of pNF-κB protein expression in astrocytes. (F) IPC+OGD enhanced the release of IFNβ compared to levels in the OGD group. (G) IPC+OGD decreased IL-6 release compared to levels in the OGD group. Values are expressed as mean ± SD of three independent experiments, each carried out in triplicate; * p <0.05, ** p <0.01.

Article Snippet: For double staining of TLR3 and glial fibrillary acidic protein (GFAP), the sections were incubated together with goat anti-TLR3 antibody (1∶50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and mouse anti-GFAP antibody (1∶200; Epitomics, Cambridge, UK) overnight at 4–8°C, and then incubated sequentially with fluorescein-labeled secondary antibody for 2 h at room temperature.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: PLoS ONE

Article Title: Astrocytic Toll-Like Receptor 3 Is Associated with Ischemic Preconditioning- Induced Protection against Brain Ischemia in Rodents

doi: 10.1371/journal.pone.0099526

Figure Lengend Snippet: (A–C) Astrocyte cultures were treated with 10 µg/mL Poly I:C plus non-immune IgG or 50 ng/mL anti-TLR3 antibody (Ab-TLR3) 24 h before being subjected to 12-h oxygen-glucose deprivation (OGD). (D–F) Astrocyte cultures were exposed to IPC and treated with non-immune IgG or 50 ng/mL Ab-TLR3 24 h before being subjected to 12-h OGD. Pretreatment of astrocyte cultures with the neutralizing antibody against TLR3 reversed IPC- and Poly I:C-induced ischemic protection and the increase in IFNβ. Values are expressed as mean ± SD of three independent experiments, each carried out in triplicate; * p <0.05, ** p <0.01, NS = not significant; LDH = lactate dehydrogenase.

Article Snippet: For double staining of TLR3 and glial fibrillary acidic protein (GFAP), the sections were incubated together with goat anti-TLR3 antibody (1∶50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and mouse anti-GFAP antibody (1∶200; Epitomics, Cambridge, UK) overnight at 4–8°C, and then incubated sequentially with fluorescein-labeled secondary antibody for 2 h at room temperature.

Techniques: