GCK RNA interference leads to cell death and cell-cycle arrest of DLBCL cell lines. (A) DLBCL cells were infected with lentiviral vectors expressing both GFP and either shRNA#1 directed against GCK or a nontargeting (control) vector. The percentage of live cells in the GCK shRNA vector expressing cells at 72 hours postinfection are presented as a percentage of the control cells. The efficacy of these shRNAs is demonstrated by immunoblotting for V5-tagged GCK in 293T cells. (B) Cell-cycle analysis of GCK shRNA#1 vector or control infected cells, representative of results in 3 independent experiments. (C) DLBCL cells were infected with lentiviral vectors with a puromycin resistance cassette and either shRNA#2 directed against GCK or a nontargeting (control) vector. ATP turnover was measured using the ATPlite System, as described in “Methods,” to simultaneously assess growth and viability. A western blot showing representative knockdown in SU-DHL-6 cells is to the right. This experiment was performed 3 times. (D) ON-TARGETplus siRNA was transfected using AMAXA electroporation to knock down GCK expression in OCI-LY-10, OCI-LY-19, SU-DHL-6, or G452 cells. Survival of cells at 72 hours posttransfection was analyzed by flow cytometry. Western blots showing representative knockdown in the same cell line are beneath each survival graph. All experiments were performed at least 3 times. Data in (A,C-D) are represented as mean ± standard error of the mean.