SHKBP1 KO enhances Nrf2 nuclear translocation. (A) IF analysis of endogenous Nrf2 in WT and SHKBP1 KO HeLa cells by confocal microscopy. Nuclei were stained with DAPI (magenta). Cells were treated with As(III) (10 µM) for the indicated times. Scale bars: 15 μm. (B) Quantification of nuclear levels of Nrf2 in WT and SHKBP1 KO HeLa cells with As(III) treatment for the indicated times (representative images shown in (A)). n = 9 images each including ∼30 cells from three independently plated samples. Exact P values indicated (****P < 0.0001) from two-way ANOVA with Tukey’s post hoc test. (C) Western blot analysis of cytosolic and nuclear fractions from WT and SHKBP1 KO HeLa cells treated with As(III) (10 µM) for the indicated times. Tubulin and Histone H3 are used to qualitatively assess the purity of each fraction. (D) Quantification of nuclear Nrf2 protein levels, with band intensities normalized to Histone H3 levels (n = 3). Exact P values indicated from two-way ANOVA. (E) Role of SHKBP1 in the p62-Keap1-Nrf2 pathway. Source data are available for this figure: SourceData F7.