The matrix component biglycan
is proinflammatory and signals through
Toll-like receptors 4 and 2 in macrophages
J. Clin. Invest. Liliana Schaefer, et al. 115:2223 doi:10.1172/JCI23755 [
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Figure 3LPS-stimulated macrophages secrete IL-6 and IL-1β, both of which induce the expression of BGN, which in turn stimulates TNF-α and MIP-2 mRNA and protein expression in macrophages. (
A and
B) ELISA for IL-6 (
A) and IL-1β (
B) in culture media from
Bgn+/0 and
Bgn–/0 macrophages left unstimulated or stimulated with 0.5 ng/ml LPS for 1 hour. (
C and
D) RT-PCR of BGN mRNA (
C) and Western blot of BGN core protein (
D) secreted into culture media from
Bgn+/0 and
Bgn–/0 macrophages 2 hours after stimulation with either IL-6 or IL-1β (both 10 ng/ml), normalized to GAPDH or β-tubulin, respectively. (
E) Northern blots of TNF-α and MIP-2 mRNA (normalized to α-tubulin) in
Bgn+/0 and
Bgn–/0 macrophages after 6 hours of incubation with BGN (4 μg/ml). (
F and
G) Dose-dependent enhancement of TNF-α (
F) and MIP-2 (
G) concentrations in media from
Bgn+/0 or
Bgn–/0 macrophages cultured for 24 hours in the absence or presence of BGN (1 or 10 μg/ml). (
H) Time-dependent enhancement of TNF-α concentrations in media from
Bgn+/0 or
Bgn–/0 macrophages cultured for 6 and 24 hours in the absence or presence of BGN (10 μg/ml). (
I and
J) ELISA for TNF-α (
I) and MIP-2 (
J) in media from
Bgn+/0 or
Bgn–/0 macrophages cultured for 6 hours in the absence or presence of LPS (0.5 ng/ml). Data are given as means ± SD from 3–7 animals. *
P < 0.05 for macrophages with versus without BGN or LPS, respectively.