Journal: PLoS ONE

Article Title: Interleukin-19 Acts as a Negative Autocrine Regulator of Activated Microglia

doi: 10.1371/journal.pone.0118640

Figure Lengend Snippet: (A) Western blotting data for receptors of IL-19. Only microglia express both the IL-20Rα and IL-20Rβ subunits. Neu, neurons; Ast, astrocytes; Mi, microglia. (B) Western blotting data for receptors of IL-19. LPS stimulation did not change the expression levels of IL-19 receptor subunits. Skin was used as the positive control. NT, untreated; LPS, LPS-treated. (C) Western-blotting analysis of STAT3 phosphorylation in microglia treated with 100 ng/ml IL-19. IL-19 induced STAT3 phosphorylation, the main downstream signal of the IL-19 receptor. All quantitative data are expressed as means ± SD (n = 3). *, p < 0.01 vs . 0 min. **, p < 0.001 vs . 0 min.

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween-20 for 1 h at room temperature, and then incubated overnight at 4°C with rabbit anti-IL-20Rα polyclonal antibodies (Merck Millipore, Billerica, MA, USA), rat anti-IL-20Rβ monoclonal antibody (eBioscience, San Diego, CA, USA), mouse anti-STAT3 monoclonal antibody (BD Pharmingen, Franklin Lakes, NJ, USA), rabbit anti-pSTAT3 polyclonal antibodies (Cell Signaling Technology, Danvers, MA, USA), or mouse anti-β-actin monoclonal antibody (Sigma-Aldrich), followed by incubation with horseradish peroxidase—conjugated secondary antibodies (GE Healthcare, Buckingham, UK) for 1 h at room temperature.

Techniques: Western Blot, Expressing, Positive Control, Phospho-proteomics