Fig. 7.

FUBP1 Is Required for Nrf2 Protein Induction and Nrf2 5′UTR Activation. FUBP1 siRNA or negative control was introduced to HeLa cells at 72 hours prior to H₂O₂ treatment. Cells were harvested at 1 hour after 10 min of 100 μM H₂O₂ treatment, for Western blot to detect FUBP1 or Nrf2 protein from total cell lysates using GAPDH as a loading control (A, B). FUBP1 siRNA or negative control was cotransfected with pRF bicistronic vector of Nrf2 5′UTR in HeLa cells. At 72 hours after, cells were treated with H₂O₂ for measurements of luciferases at 1 hour after (C). The results of one experiment representative of three are shown (A, C), or are summarized as average ± SD of Nrf2 band intensity over that of GAPDH from three experiments (B). * indicates significant difference between control versus the H₂O₂ treated group (p < 0.05), whereas # indicates significant difference between two H₂O₂ treated groups with FUBP1 siRNA versus with control siRNA (p < 0.05).