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. 2012 Jun 27;7(6):e39967. doi: 10.1371/journal.pone.0039967

Figure 8. DOK3-deficient macrophages are more sensitive to endotoxin tolerance.

Figure 8

Tolerance was induced in wild type and DOK3-deficient BMM with the indicated dose of LPS (10 or 100 ng/ml) overnight, followed by resting in fresh media for 2 h and re-challenged with the same dose of LPS for 12 h in the presence of Brefeldin A. Intracellular TNFα production was assessed with flow cytometry after the cells were fixed, permeabilized, and stained with anti-TNFα. Results are represented (A) by one-parameter histograms (green is control staining, red is naïve cells, and blue is tolerant cells) or (B) by MFI (mean fluorescence intensity) of TNFα-producing population and representative of two independent experiments. (C) Tolerant wild type and DOK3-deficient BMM or naïve cells were stimulated with 1 µg/ml LPS and immunoblotted for SHIP1, IRAK-M, SOCS1, and GAPDH. Densitometry of the bands was determined using image J normalizing to GAPDH, and the relative numerical value was written below the band. Data are representative of three independent experiments.