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. 2016 Nov 22;17(11):1938. doi: 10.3390/ijms17111938

Figure 5.

Figure 5

Figure 5

Effects of CTB on the upstream pathway of activator protein (AP)-1. RAW264.7 cells were pre-treated with 50 µM CTB for 30 min and then stimulated with 1 µg/mL LPS for the indicated time. After immunoblotting, the levels of phospho- or total forms of ERK, JNK and p38 were identified based on their antibodies (A); RAW264.7 cells were pre-treated with 10 µM p38 MAPK inhibitor (SB203580) for 30 min before treatment with 1 µg/mL LPS for 6 h. mRNA expression of iNOS, COX-2, and TNF-α was measured by real-time PCR (B); RAW264.7 cells co-transfected with plasmids containing AP-1-luc luciferase construct were treated with CTB in the presence or absence of LPS (C); Values are the mean ± SD of triplicate experiments. * p < 0.05 compared to LPS treatment alone; # p < 0.05 compared to control group. RAW264.7 cells were pre-treated with different concentrations of SB203580 for 30 min and then stimulated with 1 µg/mL LPS for 1 h. Cell lysates were immunoblotted with phospho- or total MKK3/6 (D); Superposition of the crystal structures of MKK3 and MKK6 with the docking structure of the CTB (E). A typical experiment of three independent experiments is shown.