Molecules and Cells

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Fig. 1.

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Fig. 1. (A) RAW 264.7 cells were incubated with gAd (1, 3, 5, 10 μg/ml) and LPS (1 μg/ml) for 30 min. Cells were stimulated with gAd (5 μg/ml) for the indicated times. (B) RAW 264.7 cells were pretreated with polymyxin B (PMB, 10 μg/ml) for 1 h, and then stimulated with gAd (5 μg/ml) for 30 min in the presence or absence of PMB. Cells were treated with gAd and heated gAd (5 μg/ml) for 30 min. Total cellular extracts were subjected to Western blot analysis for p-IκBα, IκBα and actin. (C) Cells were stimulated with gAd (5 μg/ml) for the indicated times. Cytoplasmic (C) and nuclear (N) proteins were extracted. Equal amounts of protein were subjected to Western blot analysis for p65. Ratio of nuclear to cytoplasmic (N/C) p65 densitometry values (lower panel of (C)). (D) RAW 264.7 cells were stimulated with gAd (5 μg/ml) for the indicated times. Total RNA was isolated and quantitative real-time PCR for TNF-α and GAPDH was performed. Data represent the mean ± SD of triplicates. **P < 0.05 (E) Cells were treated with gAd (5 μg/ml) and LPS (1 μg/ml) for 24 h. TNF-α concentration in culture medium was measured by multiplex bead assay. Data represent the mean ± SD of triplicates. **P < 0.05 Results are representative of three separate experiments. n.s., non-specific.
Mol. Cells 2018;41:762~770 https://doi.org/10.14348/molcells.2018.0005
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