Berbamine inhibits CaMKII γ-dependent NF-κB, β-catenin, and Stat3 without affecting its upstream target PKCα. (A) Effect of berbamine on PKCα, CaMKIIγ, IKK-α, and IkBα. (B-C) Berbamine analogs blocked nuclear translocation of NF-κB p65 realed by Western blot (B) and immunofluorescent staining (C). (C) Immunofluorescent image results exhibited a significant decrease of nuclear NF-κB p65 of leukemia cells after treatment of berbamine. Cells were immobilized on coverslips, fixed, permeabilized, and subjected to immunostaining of p65 (red fluorescence) and nuclear counterstaining (blue). Scale bars, 10 μm. (D-E) Berbamine inhibited CaMKIIγ downstream targetsβ-catenin (D) and Stat3 (E) of leukemia cells. Leukemia cells were treated with berbamine at various concentrations for 48 hours and harvested for analysis of upstream (PKCα) and downstream (IKK-α, IkBα,NF-κB p65, β-catenin and Stat3) targets of CaMKIIγ using Western blotting or immunofluorescent staining.