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. 2015 Dec 3;5:17642. doi: 10.1038/srep17642

Figure 6. Az produces inflammatory cytokines in cells.

Figure 6

(a) RAW264.7 cells were treated with 3 μM Az and the supernatant was collected at the indicated times. The amounts of cytokines were determined by using a multiplex cytokine assay. (b) RAW264.7 cells were treated with Az (3 μM), Alum (4 μg/ml) or PolyI:C (5 μg/ml) and supernatant was collected at the indicated times. The amount of TNF-α was determined by using an ELISA. (c) RAW264.7 cells were treated with various concentrations (0.01, 0.1, 0.2, 1 and 3 μM) of Az and the supernatant was collected at the indicated times. The amount of TNF-α was analyzed by using an ELISA. (d) RAW264.7 cells were treated with 3 μM Az and then fractionated into the cytosol and nucleus, which both were subjected to western blot analysis to detect IκBα, p-IκBα and NF-κB-p65. α-Tubulin was used as a loading control. (e) HUVEC cells transfected with a NF-κB luciferase reporter were treated with 3 μM Az or 1 μg/ml LPS for 12 h and then luciferase activity was measured. All graphs are shown as mean ± s.e.m. *p < 0.05; **p < 0.01; ***p < 0.001.