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. 2005 Jul 14;115(8):2223–2233. doi: 10.1172/JCI23755

Figure 6.

Figure 6

Absence of BGN-mediated stimulation of ERK, TNF-α, and MIP-2 in TLR2–/–/TLR4-M macrophages and reduced stimulation in macrophages from TLR2–/–, TLR4-M, and MyD88–/– mice. (A) Phosphorylation of ERK in C57BL/6, TLR2–/–, TLR4-M, TLR2–/–/TLR4-M, and MyD88–/– macrophages 30 minutes after incubation with BGN (4 μg/ml), LPS (2 ng/ml), or peptidoglycan (Pepti) (5 μg/ml) (Western blots). (B) Densitometric quantification of p-ERK/ERK ratio in macrophages indicated in A. Controls represent macrophages without BGN and are defined as 1 arbitrary unit. Data are given as means ± SD from 3 Western blots. (C and D) ELISA for TNF-α (C) and MIP-2 (D) in media from macrophages indicated in A after 6 hours of culture without or with BGN (4 μg/ml). Data are given as means ± SD from 7–8 animals in each group. Significant differences for TLR- or MyD88-deficient versus C57BL/6 macrophages incubated with BGN are indicated by an asterisk positioned over the respective bar: *P < 0.05. (E) BGN- or LPS-mediated (2 ng/ml) activation of NF-κB, detected after 30 minutes by EMSA in HEK-Blue-4 cells (transfected with human TLR4/MD-2/CD14 genes) versus 293/null cells. C1 (macrophages with 500 times free oligonucleotides) and C2 (without nuclear extract) represent negative controls.