Figure 3

Impaired activation after treatment with LPS or TNFα in Ymer Tg mice. (A) Thioglycollate-induced peritoneal macrophages isolated from WT and Ymer Tg mice were stimulated with LPS (1 ng/mL) for the indicated times, and the cell lysates were analyzed by immunoblotting to detect the activation of IKK and MAPK. The percentages of isolated macrophages were >85%. (B) Cytokine production in thioglycollate-induced peritoneal macrophages from Ymer Tg mice after treatment with LPS. mRNA expression levels of cytokines (IL-6, TNFα and IL-1 β) were analyzed by reverse transcription-polymerase chain reactions (RT-PCRs). (C) IL-6 production by thioglycollate-induced peritoneal macrophages from Ymer Tg mice after treatment with LPS. ELISA was performed to measure IL-6 production in the culture supernatant at the indicated times. (D) MEFs isolated from WT and Ymer Tg mice were stimulated with TNFα (10 ng/mL) for the indicated times, and the cell lysates were analyzed by immunoblotting to detect the activation of IKK and JNK. (E) NF-κB luciferase reporter assay using MEFs from Ymer Tg mice. Eight hours after TNFα (10 ng/mL) stimulation, NF-κB-dependent transcriptional activities were measured.