Inhibition of GSK-3 decreased NFκB-mediated expression of antiapoptotic molecules in CLL cells. (A) CLL B cells from patients with CLL were treated with diluent (DMSO) or 25 μmol/L AR-A014418; 12 hours after treatment, the cell pellet was collected and RNA was obtained. RT-PCR analysis was performed as described in “Materials and methods.” P = patient; B2M = β-2-microglobulin. (B) MEC1 cells were treated with diluent (DMSO), 25 μmol/L AR-A014418 (ARA), 50 μmol/L Z-VAD-FMK (ZVAD), or AR-A014418 + Z-VAD-FMK; the cell pellet was collected and RNA (12 hours after treatment) and protein (24 hours after treatment) were obtained. Cell lysates were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with the indicated antibodies (left panel). RT-PCR analysis was performed as described in “Materials and methods.” (C) CLL B cells from a patient with CLL were treated with diluent (DMSO) or 25 μmol/L AR-A014418 (ARA), 50 μmol/L Z-VAD-FMK (ZVAD), or AR-A014418 + Z-VAD-FMK; the cell pellet was collected and RNA (12 hours after treatment) and protein (24 hours after treatment) were obtained. mRNA and protein expression analysis was performed as described in panel B.