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. 2017 Mar 9;292(16):6644–6656. doi: 10.1074/jbc.M116.762021

Figure 4.

Figure 4.

EBI3 activates STAT3 phosphorylation in B9 cells, and this phosphorylation is IL-6-dependent. A, B9 cells were stimulated with mEBI3 (1 μg/ml), IL-27 (100 ng/ml), or IL-6 (100 ng/ml) for 15 min or 16 h with or without anti-IL-6 mAb. B, B9 cells stably transfected with mouse IL-27Rα cDNA were stimulated with IL-27 (100 ng/ml) for 15 min with or without anti-IL-6 mAb. The cells were fixed, permeabilized, and stained with FITC-labeled anti-pSTAT3. Fluorescence was assessed by flow cytometry. The filled histograms indicate unstimulated cells. Mean fluorescence intensity values of the unstimulated (upper left corners) and stimulated cells (upper right corners) are indicated in the histograms. C, B9 cells were stimulated with mEBI3 (0.5, 1, or 2 μg/ml) for 16 h, IL-27 (100 ng/ml), or IL-6 (100 ng/ml) for 15 min with or without anti-IL-6 mAb. D, B9 cells stably transfected with mouse IL-27Rα cDNA were stimulated with IL-27 (100 ng/ml) for 15 min with or without anti-IL-6 mAb. Cell lysates were analyzed by Western blot using anti-pSTAT3 or anti-STAT3 followed by anti-rabbit IgG-HRP. The data are representative of three independent experiments.