FIG. 6.

Negative regulation of cytokine response by TLR-mediated endotoxin tolerance. (A to D) Supernatants from MC treated with LPS as described in Materials and Methods were analyzed for TNF-α (A), IL-8 (B), IL-6 (C), and IL-10 (D) by flow cytometry using a CBA kit. The assay was performed in triplicate. Based on the standard curve for each cytokine, the CBA software calculates levels in picograms per milliliter. (E) Mean levels of IL-1β mRNA from triplicate samples analyzed by real-time PCR, as described in the legend to Fig. 3 and in Materials and Methods. Error bars indicate standard deviation.