Src Is a Prime Target Inhibited by Celtis choseniana Methanol Extract in Its Anti-Inflammatory Action

Figure 3

The effect of Cc-ME on activation of the NF-κB signaling pathway. (a, b, and c) RAW264.7 cells pretreated with Cc-ME (100 μg/ml) for 30 min were treated with LPS (1 μg/ml) for the indicated time, and phosphorylated and total forms of Src, p85, AKT, IKKα/β, IκBα, and β-actin were examined by Western blot analysis using their specific antibodies. (d) HEK293 cells were transfected with HA-Src for 48 h, and phosphorylated and total forms of Src, p85, AKT, IKKα/β, IκBα, and β-actin were examined by Western blot analysis using their specific antibodies. (e) RAW264.7 cells pretreated with Cc-ME (100 μg/ml) for 30 min were treated with LPS (1 μg/ml) for 5 min. Phosphorylated Src was immunoprecipitated in the total cell lysates of the cells, and phosphorylated p85 was examined by Western blot analysis using its specific antibody. The data presented in (a), (b), (c), (d), and (e) are a representative of three experiments. Relative intensity (right panels of (a), (c), and (d), and bottom panel of (e)) was expressed as means ± SD, calculated with data observed by three experiments using the DNR Bioimaging system. WCL: whole cell lysate. HC: heavy chain. and compared to control group.

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