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. 2015 Dec 14;212(13):2289–2304. doi: 10.1084/jem.20150388

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© 2015 Phong et al.

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Figure 5. — Nur77-GFP reporter expression is induced by IgE/Ag and enhanced by coengagement of Tim-3. BMMCs were generated from Nur77^GFP reporter mice, sensitized with IgE overnight, and stimulated with Ag. In A, Ag (DNP₃₂-HSA) was titrated over a range of 10–500 ng/ml. PMA plus ionomycin stimulation was used as a positive control. GFP expression was determined after stimulation for 6 h. (B) Nur77^GFP BMMCs were stimulated with a fixed concentration of Ag, with or without Tim-3 pAb for 6 h. (C) Nur77^GFP BMMCs were stimulated with IgE/Ag and either Tim-3 pAb or Tim4-Fc (which binds to Tim-1), or appropriate isotype controls. (D) Nur77^GFP BMMCs were stimulated with IgE/Ag plus either inhibitor to Src kinases (PP2) or Syk (BAY). (E) Nur77^GFP BMMCs were stimulated with IgE/Ag and the indicated inhibitors to MEK (UO126), Akt (Akti1/2), calcineurin (CsA), PI3K (LY294002), or PKC (BIM). GFP signal was quantified by flow cytometry. Results are the average of three independent experiments performed in duplicate. *, P < 0.05; **, P < 0.005; ***, P < 0.0005.