Fig 1: 10v induces the activation and nuclear translocation of Nrf2. (A) Concentration–response curve of 10v in the Keap1-Nrf2 nuclear translocation assay using engineered U2OS cells. Nrf2 activation was quantified by measuring relative light units (RLU) emitted by Nrf2 translocation. (B) Cell viability in BV-2 microglial cells after 24 h of 10v treatment. (C) Western blot analysis of Nrf2 in the nuclear fraction of BV-2 microglial cells following time-dependent treatment with 10v. (D) Relative protein levels of Nrf2 normalized to Lamin B1. (E,F) Nrf2 accumulation after 6 h of 10v treatment in whole cell lysates (E) and nuclear fraction (F) of BV-2 microglial cells. All experiments were repeated at least twice. Immunoblotting images are representative results of repeated experiments, and relative fold expressions were determined based on the intensity of each band. Data are represented as mean ± SEM. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, One-way ANOVA with Dunnett’s test compared to vehicle-treated control.
Fig 2: 10v induces Nrf2-dependent antioxidant gene expression. (A,B) Relative expression of Hmox1 (A) and Gclm mRNA (B) after 12 h of 10v treatment in BV-2 microglial cells. Relative expression levels were normalized to Hprt mRNA. (C) Western blot analysis of HO-1 and GCLM after 24 h of 10v treatment in BV-2 microglial cells. (D,E) Relative protein levels of HO-1 (D) and GCLM (E) normalized to β-actin. All experiments were repeated at least twice. Immunoblotting images are representative results of repeated experiments, and relative fold expressions were determined based on the intensity of each band. Data are represented as mean ± SEM. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, One-way ANOVA with Dunnett’s test compared to vehicle-treated control.
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