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. 2020 Feb 4;17(4):498–509. doi: 10.7150/ijms.37833

Figure 3.

Figure 3

Figure 3

Figure 3

S100A8 and S100A9 require activation of PI3K/AKT/ MAPKs/NF-κB pathway for cytokine expression. (A) BEAS-2B cells were incubated with 10 μg/mL S100A8 (left panel) and S100A9 (right panel) for the indicated time. Phosphorylation of AKT, ERK, p38 MAPK and JNK in the lysates was detected by Western blotting. (B) BEAS-2B cells were pre-treated for 1 h with or without 5 μM TLR4i, 5 μM LY294002 (LY) and 10 μM AKTi, after which the cells were incubated with 10 μg/mL S100A8 (left panel) and S100A9 (right panel) for 30 min. After harvested cells were lysed, the phospho-ERK (p-ERK), phospho-p38 MAPK (p-p38 MAPK) and phospho-JNK (p-JNK) in the lysates were detected by Western blotting. Densitometric data are presented relative to the negative control set at l (lower panel). (C, D) BEAS-2B cells were incubated with S100A8 and S100A9 (10 μg/mL) for the indicated time (C), or were pre-treated for 1 h with or without 5 μM TLR4i, 5 μM Ly294002 (LY) and 10 μM AKTi, 10 μM PD98059 (PD), 10 μM p38 MAPK (SB) and 10 μM JNK600125 (SP), after which the cells were incubated with S100A8 and S100A9 (10 μg/mL) for 1 h (D). After collecting nuclear extract from harvested cells, NF-κB in the lysates (n=3) was detected by luciferase assay. Data are presented relative to the control set at 1, as the means ± SD. *p < 0.05 and **p < 0.01 indicate a significant difference between the control and S100A8 and S100A9-treated groups, and ##p < 0.01 and #p < 0.05 represent a significant difference between the S100A8- or S100A9-treated and inhibitor-treated groups.