Mediators of Inflammation / 2019 / Article / Fig 3

Research Article

Electronegative LDL from Rabbits Fed with Atherogenic Diet Is Highly Proinflammatory

Figure 3

LDL(-)-induced activation of NF-κB and expressions of NF-κB downstream genes. (a) THP-1 macrophages were incubated with 20 μg/ml of native (n)LDL or LDL(-) for 2 h, and then protein levels of total and phosphorylated IκB (IκB and p-IκB, respectively) and p65 (p65 and p-p65, respectively) were determined by Western blotting. β-Actin was used as a loading control. (b, c) Relative levels of p-IκB/total IκB (b) and p-p65/total p65 (c) were expressed relative to the control (PBS, ). Values are the of three independent experiments. , compared to PBS- and nLDL-treated cells. (d) THP-1 macrophages were treated with 20 μg/ml of nLDL or LDL(-) for 6 h, and levels of IL-1β, IL-6, TNF-α, CD86, and IL-10 mRNAs were determined by an RT-qPCR, normalized to levels of GAPDH mRNA, and expressed relative to levels in nLDL-treated cells (). , compared to nLDL-treated cells. (e–g) Cells were pretreated with 10 μM BAY 11-7082 for 1 h and then treated with LDL(-) (20 μg/ml) for 24 h. Levels of IL-1β (e), IL-6 (f), and TNF-α (g) in the medium were determined by ELISA. , compared to DMSO-treated cells.

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