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. 2010 Nov 2;29(24):4223–4236. doi: 10.1038/emboj.2010.268

Figure 1.

Figure 1

FoxO1 enhances Tlr4 signalling in macrophages. (A) RAW264.7 cells were infected with either Ad-GFP or Ad-FoxO1-CA at 100 MOI. After 48 h, cells were exposed to 100 ng/ml LPS for 30 min and then lysed for immunoblotting with the indicated antibodies. (B) Relative amount of phosphorylated versus total protein levels for P65 and JNK were quantitated by NIH-Image J; and the results are shown as average±s.d. (C, D) RAW264.7 cells were co-transfected with FoxO1-WT, FoxO1-CA or DBD mutant FoxO1 plasmid vectors together with NFκB-Luc (C) or iNOS-Luc (D), and then exposed to 100 ng/ml LPS or PBS control for 6 h before luciferase assay. (E) RAW264.7 cells were stably transduced with lentivirus encoding shRNA against luciferase (control) or FoxO1, then exposed to 100 ng/ml LPS for 15 and 30 min, and subsequently lysed for immunoblotting assays with indicated antibodies. (F) RAW264.7 cells were stably transduced with lentivirus encoding shRNA against luciferase or FoxO1 and then exposed to 100 ng/ml LPS for 6 h. iNOS mRNA expressions levels were assayed by real-time PCR. Data are presented as the average±s.d. Letters above the bars show statistical groups (ANOVA, P<0.05).