rabbit polyclonal against lysozyme antibody  (AnaSpec)

Journal: Frontiers in Molecular Neuroscience

Article Title: Hypoxic Preconditioning Maintains GLT-1 Against Transient Global Cerebral Ischemia Through Upregulating Cx43 and Inhibiting c-Src

doi: 10.3389/fnmol.2018.00344

Figure Lengend Snippet: Cellular localization of Cx43 in CA1 in Sham group and tGCI group with or without hypoxia at 168 h of reperfusion. Representative images of triple fluorescent staining of Cx43 (green), NeuN (red) and DAPI (blue); Cx43 (green), GFAP (red) and DAPI (blue); Cx43 (green), Iba-1 (red) and DAPI (blue) in CA1. (A) The overlapped images showed that Cx43 prominently surrounded NeuN (a–d) , and located slightly in GFAP-positive (e–h) and Iba-1-positive (i–l) cells in Sham group. (B) Cx43 mainly located in GFAP-positive (e–h) and Iba-1-positive (i–l) cells at 168 h after tGCI, and part of cells with NeuN (a–d) . (C) Cx43 located mainly in GFAP-positive (e–h) and Iba-1-positive (i–l) , and partly in NeuN-positive cells (a–d) in CA1 at 168 h after reperfusion of HPC group. Scale bar: 25 μm. (D–F) Quantitative analysis of Cx43 positive neurons, Cx43 positive astrocytes and Cx43 positive microglia in the CA1, respectively. Data are expressed as percentage of the number of Cx43-positive cells. Each bar represents the mean ± SD; * p < 0.05 vs. Sham group and # p < 0.05 vs. tGCI group ( n = 4 in each group). Sham, sham-operated; tGCI, transient global cerebral ischemia; HPC, hypoxic preconditioning; Cx43, connexin 43; NeuN, neuronal nuclei; GFAP, glial fibrillary acidic protein; Iba-1, ionized calcium binding adaptor molecule 1; DAPI, 4’,6-diamidino-2-phenylindole.

Article Snippet: Sections were preincubated with 5% normal serum (containing 0.2% Triton X-100) for 1 h at room temperature, and then incubated overnight at 4°C with mixtures of primary antibodies: goat polyclonal antibody against Cx43 (1:100; Abcam, Cat# ab219493), mouse monoclonal antibody against NeuN (1:1,000; Millipore, Cat# MAB377, RRID: AB_2298772 ), rabbit polyclonal antibody against Iba-1 (1:100, Abcam, Cat# ab108539, RRID: AB_10862652 ).

Techniques: Staining, Binding Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Hypoxic Preconditioning Maintains GLT-1 Against Transient Global Cerebral Ischemia Through Upregulating Cx43 and Inhibiting c-Src

doi: 10.3389/fnmol.2018.00344

Figure Lengend Snippet: Cellular localization of p-c-Src in CA1 in Sham group and tGCI group with or without hypoxia at 168 h of reperfusion. Representative images of triple fluorescent staining of p-c-Src (green), NeuN (red) and DAPI (blue); p-c-Src (green), GFAP (red) and DAPI (blue); p-c-Src (green), Iba-1 (red) and DAPI (blue) in CA1. (A) The overlapped images showed that p-c-Src was mainly colocalized with NeuN positive cells (a–d) , rather than GFAP (e–h) and Iba-1 (i–l) positive cells in Sham group. (B) p-c-Src mainly located in Iba-1-positive (i–l) cells at 168 h after tGCI, and a minority of p-c-Src positive cells were NeuN-positive (a–d) , rather than GFAP-positive (e–h) . (C) p-c-Src located mainly in NeuN-positive (a–d) and Iba-1 positive (i–l) cells, but not in GFAP-positive (e–h) cells in CA1 at 168 h after reperfusion of HPC group. Scale bar: 25 μm. (D–F) Quantitative analysis of p-c-Src positive neurons, p-c-Src positive astrocytes and p-c-Src positive microglia in CA1, respectively ( n = 4 in each group). Data are expressed as percentage of the number of p-c-Src-positive cells. Each bar represents the mean ± SD * p < 0.05 vs. Sham group and # p < 0.05 vs. tGCI group. Sham, sham-operated; tGCI, transient global cerebral ischemia; HPC, hypoxic preconditioning; p-c-Src, phosphorylated cellular-Src; NeuN, neuronal nuclei; GFAP, glial fibrillary acidic protein; Iba-1, ionized calcium binding adaptor molecule 1; DAPI, 4’,6-diamidino-2-phenylindole.

Article Snippet: Sections were preincubated with 5% normal serum (containing 0.2% Triton X-100) for 1 h at room temperature, and then incubated overnight at 4°C with mixtures of primary antibodies: goat polyclonal antibody against Cx43 (1:100; Abcam, Cat# ab219493), mouse monoclonal antibody against NeuN (1:1,000; Millipore, Cat# MAB377, RRID: AB_2298772 ), rabbit polyclonal antibody against Iba-1 (1:100, Abcam, Cat# ab108539, RRID: AB_10862652 ).

Techniques: Staining, Binding Assay