FIG. 2.

The TLR5 ligand flagellin elicits transcriptional factor activation and inflammatory cytokine production in murine osteoblasts. (A) Cells were unexposed or exposed to purified flagellin (10 and 100 ng/ml) for 90 min. Whole-cell lysates were analyzed by Western blotting for the presence of phosphorylated JNK1/2. (B) Cells were unexposed or exposed to purified flagellin (0.1, 1, 10, and 100 ng/ml) for 7 h prior to RNA isolation and RT-PCR for mRNA encoding IL-6. These studies were performed three times with similar results. (C) Cells were unexposed or exposed to purified flagellin (1, 10, and 100 ng/ml) for 24 h. Culture supernates were analyzed for the presence of IL-6 protein by specific capture ELISA. Results shown are the means ± standard errors of the means of replicate measurements in four separate preparations. *, significantly different from untreated cells (P < 0.05) when tested by Student's paired t test.