Figure 7.

The Super LKB1+ cells presented a cytoplasmatic localization of NRF2 and lower expression of antioxidant enzymes.A, Western blotting of NRF2, KEAP1, NQO1, PRDX3, and β-Actin in cWT, c2SL+, and c3SL + cells; (B) normalized levels of proteins by β-Actin (a.u); (C) immunofluorescence of NRF2 in cWT, c2SL+, and c3SL + cells; (D) subcellular fractionation of nucleus and cytosolic, followed by Western Blotting of NRF2, KEAP1, and β-Actin cWT, c2SL+, and c3SL + cells; (E) plot profile analysis of fluorescence distance in cWT cells, the green line refers to NRF2, and the blue line refers to the nucleus stained with DAPI. F, plot profile analysis of fluorescence distance in c2SL + cells, the green line refers to NRF2, and the blue line refers to the nucleus stained with DAPI. G, plot profile analysis of fluorescence distance in c2SL3+ cells, the green line refers to NRF2, and the blue line refers to the nucleus stained with DAPI. Red arrows in plot profiles represent peaks of signal colocalization. H, nuclear fluorescence quantification in cWT, c2SL+, and c3SL+. I, cytosolic fluorescence quantification in cWT, c2SL+, and c3SL+. J, relative H₂O₂ level (μM) in cWT and c2SL + cells in the control condition, and the treatments with metformin 10 mM and 3-MA 200 μM for 72 h. K, relative mRNA expression of NQO1, SOD1, and SOD2 in cWT, c2SL+, and c3SL + cells. The data are presented as mean ± SD. Statistical analysis was performed by ANOVA followed by Dunnet’s test ∗p < 0.05, ∗∗p < 0.01. These data are representative of two independent experiments.