Fig. 1

Mild ROS production leads to defective nuclear translocation of BACH1 via activating PI3K-AKT pathway in resistant MCL cells. a Immunoblots for BACH2 in Jeko and REC-1 cells with GAPDH as a loading control. b ROS production was measured in cells treated with DMSO (control) or BTZ (20 nM) for 24 h. Relative MFI values compared to control are shown as the mean ± SD from two independent experiments. c Cells were treated with DMSO or BTZ (20 nM) for 24 h. Cell viability was determined using MTT assays. Data are normalized to control and shown as the mean ± SD from three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 (vs DMSO-treated group). d Immunoblots for p-AKT and AKT in cells with DMSO or BTZ treatment (20 nM), with Actin as a loading control. The indicated values of p-AKT under each lane are normalized to Actin. e Cells were treated with BTZ (20 nM) with or without 1 h-pretreatment of antioxidant NAC (100 μM) for 24 h. p-AKT and AKT were measured using immunoblotting with Actin as a loading control. The normalized values (p-AKT/Actin) in each lane are indicated. f Immunoblots for BACH2 in the cytoplasmic and nuclear fractions of MCL cells treated with BTZ in the presence or absence of 3-MA (5 mM) for 24 h (left). Cells treated with DMSO were used as negative controls. Actin and TBP were used as loading controls for cytoplasmic and nuclear proteins, respectively. The % proportion of the relative BACH2 levels in cytoplasm and nucleus are indicated (right)