Journal: Journal of Neuroinflammation

Article Title: Heterogeneous induction of microglia M2a phenotype by central administration of interleukin-4

doi: 10.1186/s12974-014-0211-6

Figure Lengend Snippet: Time course of IL4-induced M2 gene expression in frontal cortex and striatum . Following the indicated time intervals after intracerebroventricular injection of saline (Veh) or IL4, the RNA extracted from the frontal cortex (Fr cortex) and striatum was analyzed by real time PCR to evaluate (A) Fizz1 , (B) Arg1 , and (C) Ym1 gene expression. Data sets for each gene were calculated using the 2 -ddCt method with respect to the mean value of the 8 h vehicle group. Bars represent mean values ± SEM. * P < 0.05 versus Veh; ** P < 0.01; *** P < 0.0005 versus Veh; ° P < 0.05 versus IL4 8 h; °° P < 0.05 versus IL4 16 h ( n = 4 to 6); a P < 0.0005 versus striatum 8 h; b P < 0.05 versus striatum 8 h; bb P < 0.05 versus striatum 16 h.

Article Snippet: For Iba1 and Arg1 immunohistochemistry, sections were immunostained with rabbit polyclonal anti-Iba1 antibody (ionized calcium-binding adapter molecule-1, 019-19741, Wako Chemicals, Richmond, VA, USA), and goat polyclonal anti-Arg1 antibody (sc-18354, Santa Cruz Biotechnology, Heidelberg, Germany); sections were than incubated with biotinylated goat anti-rabbit (Iba1) and rabbit anti-goat (Arg1) secondary antibodies (VC-BA-1000-MM15 and VC-BA-5000-MM15, Vector Laboratories, Petersborough, UK), labeled by the avidin-biotin-peroxidase procedure with a commercial immunoperoxidase kit (VECTASTAIN® Elite ABC-Peroxidase Kit Standard, VC-PK-6100-KI01, Vector Laboratories).

Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: Heterogeneous induction of microglia M2a phenotype by central administration of interleukin-4

doi: 10.1186/s12974-014-0211-6

Figure Lengend Snippet: Selective induction of Arg1 expression in a subset of microglia. Brains after 16 h intracerebroventricular treatment with vehicle (A , C) and IL4 (B , D) were formalin fixed and embedded in paraffin and analyzed by immunohistochemistry to visualize Iba1 (A , B) or Arg1 (C , D) in the frontal cortex. Arg1 expression is observed selectively in microglia-like cells after IL4 treatment and is restricted to a subset of cells (D) . Scale bar 100 μm. The results obtained by counting the number of Arg1-positive microglia cells and the total number of Iba1-positive microglia cells demonstrated that an ≈ 35% subset of microglia cells is Arg1 responder microglia (E) . Veh., vehicle.

Article Snippet: For Iba1 and Arg1 immunohistochemistry, sections were immunostained with rabbit polyclonal anti-Iba1 antibody (ionized calcium-binding adapter molecule-1, 019-19741, Wako Chemicals, Richmond, VA, USA), and goat polyclonal anti-Arg1 antibody (sc-18354, Santa Cruz Biotechnology, Heidelberg, Germany); sections were than incubated with biotinylated goat anti-rabbit (Iba1) and rabbit anti-goat (Arg1) secondary antibodies (VC-BA-1000-MM15 and VC-BA-5000-MM15, Vector Laboratories, Petersborough, UK), labeled by the avidin-biotin-peroxidase procedure with a commercial immunoperoxidase kit (VECTASTAIN® Elite ABC-Peroxidase Kit Standard, VC-PK-6100-KI01, Vector Laboratories).

Techniques: Expressing, Immunohistochemistry

Journal: Cells

Article Title: TRPA1 Expressed by Hepatocytes and Liver Macrophages Does Not Mediate Inflammatory Infiltration and Steatosis in a Mouse Model of Chronic Alcohol-Induced Liver Injury

doi: 10.3390/cells15050423

Figure Lengend Snippet: Expression of transient receptor potential ankyrin 1 ( Trpa1 , red) mRNA in the mouse liver. Trpa1 mRNA is localized both to arginase-1 (ARG, white)-positive hepatocytes ( A , A′ ) and liver macrophages co-labeled by immunostaining for CD68 (white, ( B )) and calcium-binding adaptor molecule 1 (IBA1, green, ( C )). Note the co-localization of CD68 and IBA1 in the merged images ( D , D′ ). 4′,6-diamidino-2-phenylindole (DAPI, blue) was used to visualize cell nuclei. Scale bars: 10 µm.

Article Snippet: Another set of samples was incubated with rabbit arginase-1 (1:50; Cat. No: ACI3058; Biocare Medical, Pacheco, CA, USA) primary antibody to identify the hepatocytes.

Techniques: Expressing, Labeling, Immunostaining, Binding Assay