Journal: Experimental & Molecular Medicine

Article Title: Integrin CD11b negatively regulates Mincle-induced signaling via the Lyn–SIRPα–SHP1 complex

doi: 10.1038/emm.2017.256

Figure Lengend Snippet: The Lyn activator MLR1023 inhibits TDM signaling both in vivo and in vitro . ( a ) Production of IL-6 by WT and CD11b −/− BMMs challenged with TDM and MLR1023 (1 ng ml −1 ) or DMSO for 24 h was assessed by ELISA. ( b ) Western blotting assay of Syk and Erk kinase activity and Mincle induction in WT and CD11b −/− BMMs treated with TDM and MLR1023 (1 ng ml −1 ) or DMSO for 24 h. β-Actin protein expression was used as a loading control. Experimental and control mice ( n =8 mice per group) were intravenously injected with 3.75 mg kg −1 TDM in an oil-in-water emulsion on day 0. Then the experimental mice were intraperitoneally injected with MLR1023 (6 mg kg −1 in PBS) every day beginning on day 1, until the mice were killed 7 days post-TDM challenge. The control mice were injected with 1% DMSO in PBS. ( c ) Lung tissues were isolated and stained with hematoxylin and eosin (H&E) for histology analysis after the lung weight index (LWI) ( d ) was determined. ( e ) Leukocyte subsets were analyzed by flow cytometry using distinct markers for monocytes and macrophages (Mo/Ma; CD11b+ Ly6G−), neutrophils (PMN; CD11b+ Ly6G+), T cells (CD3+) and B cells (CD19+). ( f ) Lung homogenates were analyzed by ELISA for TNF-α and IL-6. Data are representative of two independent experiments. ( c – f , mean and s.d. of eight mice per group.) * P <0.05, ** P <0.01 (two-tailed unpaired Student’s t -test).

Article Snippet: Parthenolide (0610, 5 μ M ), U0126 (1144, 10 μ M ), wortmannin (1232, 10 μ M ), Lyn peptide inhibitor (2265, 2 μ M ) and Lyn activator MLR1023 (4582, 1 ng ml −1 ) were purchased from Tocris (Ellisville, MO, USA).

Techniques: In Vivo, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay, Expressing, Control, Injection, Emulsion, Isolation, Staining, Flow Cytometry, Two Tailed Test