Treatment with LAQ824 treatment failed to induce Bak, Bim, and Bax, but attenuated Bcl-x_L and XIAP levels in HL-60/LR versus HL-60 cells. (A) After treatment of HL-60 and HL-60/LR cells with the indicated concentrations of LAQ824 for 8 and 24 hours, total cell lysates were harvested and immunoblot analyses were done for Bcl-2, Bcl-x_L, XIAP, Mcl-1, Bak, Bax, and Bim. β-actin expression served as the control for protein loading. (B) HL-60 and HL-60/LR cells were treated with the indicated concentrations of Apo-2L/TRAIL for 24 hours. After this, cell lysates were obtained and immunoblot analyses were done for DR4, DR5, caspase-8, c-FLIP, and FADD. β-actin expression in the lysates served as the loading control. (C) HL-60 and HL-60/LR cells were treated with the indicated concentrations of LAQ824 for 24 hours. Then, immunoblot analyses were done for p-AKT, AKT, p-GSK3β, p-ERK1/2, ERK1/2, c-Raf, cyclin D1, p-STAT5, STAT5, TBP-2, and TRX. The levels of β-actin expression in the lysates served as the loading control. (D) HL-60 and HL-60/LR cells were treated for 16 hours with 250 nM LAQ824. Then cells were stained with HPF and ROS levels were determined by flow cytometry. Values represent the means plus or minus SE of 3 experiments.