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. 2017 Sep 28;18(10):2069. doi: 10.3390/ijms18102069

Figure 5.

Figure 5

Effect of NNFE on nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated HO-1 expression. RAW 264.7 cells were pretreated for 24 h with NNFE. The mRNA expression (A) and protein expression (B) of Nrf2 and Kelch-like ECH-associated protein 1 (Keap1) were measured by RT-PCR and western blot, respectively. (C) The time-dependent effect on the protein levels of cytosolic and nuclear Nrf2 was analyzed by western blot. (D) Cells were treated with an Nrf2 inhibitor (brusatol) with and without NNFE, and Nrf2 and HO-1 protein levels were analyzed by western blot. (E) Cells were treated with a siRNA of Nrf2 (si-Nrf2) with and without NNFE, and Nrf2 and HO-1 protein levels were analyzed by western blot. Statistical values are expressed as the mean ± SD (n = 3). # p < 0.001, and * p < 0.05 compared to no treatment, ** p < 0.01 compared to brusatol treatment. Statistical analysis was carried out using one-way ANOVA. CA: Caffeic acid 5 μM. (−): no treatment (+): treatment with inhibitor and/or sample.