PB-020 in combination with MBZ potently reduced the level of pAKT1-S473 in CRC cells. (A) Five CRC cell lines were treated with a vehicle (CK), PB-020 (1 μM), MBZ (0.25 μM) or PB-020/MBZ combination for 72 h. Immunoblotting was conducted with the indicated primary antibodies. (B) HT-29 and LoVo cells were treated with a vehicle (CK), PB-020, MBZ or PB-020/MBZ combination for 24 h. Immunoblotting was conducted with the indicated primary antibodies. (C) All band intensities in (B) were quantified by ImageJ software and plotted as relative protein levels. (D) HT-29 cells were treated with a vehicle (CK), PB-020, MBZ or PB-020/MBZ combination for either 24 h or 48 h. Immunoblotting was conducted with the indicated primary antibodies. (E,F) All band intensities of 24 hour-treated samples in (D) were quantified by ImageJ software and plotted in (E), and all band intensities of 48 hour-treated samples in (D) were quantified by ImageJ software and plotted in (F). Significances were determined as p < 0.05 (*), p < 0.01 (**), and p < 0.001 (***).