Inhibition of NF-κB activity reduces BAFF expression and the AKT/BCL10/BCL3 signaling transduction pathway. (A) Inhibition of NF-κB activation decreases AKT activation, BCL3 nuclear translocation, and BCL10 nuclear translocation. Pfeiffer cells were treated with 20 ng/mL of BAFF for 24 hours with or without LY294002 (1-hour pretreatment as indicated). The whole-cell lysates were subjected to immunoblotting with anti-pAKT antibody, anti-BCL3 antibody, anti-BCL10 antibody, and anti–NF-κB p65 antibody. The membrane was reprobed with anti–α-tubulin antibody as a loading control. (B) Inhibition of NF-κB and BAFF activation down-regulates c-myc and cyclin D3 expression. Pfeiffer cells were treated with 20 ng/mL of BAFF for 24 hours with or without LY294002 (0-50 μM, 1-hour pretreatment as indicated). Pfeiffer cells were also treated with 20 ng/mL of BAFF for 24 hours with or without 10 μM LY294002 (1- to 24-hour pretreatment as indicated). The protein levels of c-myc and cyclin D3 were determined by immunoblotting (IB). Immunoblotting of actin served as a loading control.