Molecules and Cells

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Fig. 4.

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Fig. 4. (A, B) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for phospho-mTOR (Ser2448) (p-mTOR), sirtuin1, Nrf2, KEAP1, and GAPDH (A). Total RNA was isolated and quantitative real-time PCR for Nrf2 and GAPDH was performed (B). Data represent the mean ± SD of triplicate experiments. **P < 0.05 (C) NCI-H157 cells were pretreated with cycloheximide (CHX, a protein synthesis inhibitor, 10 μg/mL) for 1 h and then stimulated with PS-341 or MG132 in the presence or absence of CHX for 6 or 8 h. (D) NCI-H157 cells were infected with adenovirus vectors expressing wild-type Nrf2 (Ad-WT-Nrf2) or dominant-negative Nrf2 (Ad-DN-Nrf2). Forty-eight hours after infection, the cells were treated with PS-341 (50 nM) for 8 h. Cells were transiently transfected with control siRNAs or Nrf2 siRNAs. After 48 h, the cells were stimulated with PS-341 for 8 h. Total cellular extracts were subjected to Western blot analysis for Nrf2, IκBα, LC3B, and GAPDH. Results are representative of three independent experiments. (E) Densitometric analysis of the IκBα expression is shown. Results were normalized by arbitrarily setting the densitometry of non-stimulated cells to 1.0 (shown is mean ± SD; n=3). **P < 0.05
Mol. Cells 2018;41:1008~1015 https://doi.org/10.14348/molcells.2018.0277
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