Figure 5

Differentiation modified the nuclear ratio between Nrf2 and Bach1 in cell treated with H₂O₂. (a and b) Western Blotting analysis of Bach1 cytosolic and nuclear levels of undifferentiated or 4d-ATRA differentiated cells treated for 3 h with 500 µM H₂O₂ or 50 µM tBHQ, as indicated. Statistical analysis: n = 3, *p < 0.05 vs ctr. (c and d) WB analysis of Nrf2 cytosolic and nuclear levels of undifferentiated or 4d-ATRA differentiated cells treated with 500 µM H₂O₂ or 50 µM tBHQ, as indicated. Statistical analysis: n = 3, *p < 0.05 vs ctr. The expression of GAPDH and Lamin B was checked in all the experimental conditions to verify the purity of cell fractioning and then used as loading control to normalize protein expression. (e) Nrf2/Bach1 ratio has been calculated to emphasize the different behavior in the two experimental conditions. Statistical analysis: n = 3, *p < 0.05 vs ctr. Full-length blots are presented in supplementary information.