Fig. 2

BACH2 blockade leads to upregulation of HO-1 and confers BTZ-resistant properties to MCL. a Immunoblots for HO-1 in cells with or without BTZ treatment (20 nM) for 24 h. Actin was used as a loading control. b HO-1 was measured using immunoblotting in REC-1 cells treated with BTZ (20 nM) in the presence or absence of 3-MA (5 mM) for 24 h, with Actin as a loading control. c The knockdown efficiency of BACH2 (BACH2^KD) in Jeko cells was evaluated with a non-silencing shRNA plasmid (BACH2^Con) as a negative control. The basal levels of HO-1 were measured in BACH2^KD and BACH2^Con cells with Actin as a loading control. d Immunoblots for HO-1 after BTZ treatment (20 nM). Actin was used as a loading control. e ROS levels in manipulated Jeko cells with DMSO or BTZ treatment (20 nM) for 24 h. Relative MFI values are shown as the mean ± SD from two independent experiments. f Cell viability was measured in cells treated with DMSO or BTZ (20 nM) for 24 h. Data are normalized to control and shown as the mean ± SD from three independent experiments. N.S, not significant; *p < 0.05; **p < 0.01; ***p < 0.001 (vs DMSO-treated group). g Immunoblots for p-AKT and AKT in BACH2^KD and BACH2^Con cells treated with DMSO or BTZ (20 nM). The normalized values (p-AKT/Actin) in each lane are indicated. h NRF2 and LC3II were measured in manipulated Jeko cells treated with BTZ (20 nM) with or without NAC pretreatment (100 μM) for 24 h (left). The relative expressions of NRF2 (NRF2/Actin) and LC3II (LC3II/Actin) in BACH2^KD and BACH2^Con are indicated, respectively (right). Data are normalized to DMSO-treated BACH2^Con cells