Figure 4 .

NRF2 down-regulates autophagy proteins after long exposure to oxidative stress in the HEK293T cell line. A) Time dependency of autophagy down-regulation by NRF2 in oxidative stress. Western blot results of HEK293T cells in 100 µM TBHP-containing media. The Western blot shows the time course levels of autophagy-related proteins. Cells were cultured with TBHP for 0.5, 1, 2, 3, and 4 h, while NRF2 gene expression was depleted by NRF2 siRNA. GAPDH was used as a loading control. B) Quantification and statistical analysis of the Western blot assays. Densitometry data represent the intensity of p62 normalized for GAPDH, LC3-II normalized for LC3-I, ULK555-P normalized for total level of ULK, and AMPK-P normalized for GAPDH. Samples were compared with their partner with siNRF2 background. Error bars represent ± sem. *P < 0.05, **P < 0.01, ***P < 0.005 (independent 2-sample Student’s t tests).