FIGURE 3.

TLRs induce OPN expression in vivo. A, female C57BL/6J mice (4 weeks old) were intraperitoneally injected with thioglycolate to elicit peritoneal macrophages. After 3 days, the mice were treated with PBS or 1.8 mg/kg LPS intraperitoneal administration for the indicated time period. Secreted OPN and TNF-α production in the serum were detected by ELISA. B, female C57BL/6J mice (4 weeks old) were treated as in A, Secreted OPN and TNF-α production in the lavage were detected by ELISA. C, female C57BL/6J mice (4 weeks old) were treated as in A, intracellular OPN expression in the peritoneal macrophages was detected by Western blot. Actin was used as a cytoplasmic protein loading control. The expression levels were quantitated by measuring band intensities and shown as fold increase relative to background. D and E, primary splenocytes from female C57BL/6J mice (4 weeks old) were treated with 100 ng/ml LPS as indicated. Secreted OPN in supernatants was measured by ELISA. Intracellular OPN expression was detected by Western blot. Actin was used as a cytoplasmic protein loading control. Data are shown as mean ± S.D. (n = 3) of one representative experiment.