Fig. 4.

E2A knockdown alters cell cycle distribution. CLL cells were mock-transfected (no siRNA) or transfected with control siRNA or E2A siRNA, cultured without or with anti-IgM + anti-CD40 antibodies and 48 h later, cells were stained with intracellular Ki-67 and DAPI and analyzed immediately by flow cytometry. Representative staining profiles of two independent donors (CLL-6 and CLL-18) are shown in (A). DAPI and Ki-67 staining are used to visualize the G₀ compartment (lower left quadrant), G₁ compartment (upper left quadrant) and S/G₂/M compartment (upper right quadrant). Dot plots in (B) represent data across multiple independent CD38⁺ (CLL-14, -16, -18 and -22) and CD38⁻ (CLL-6, -13 and -17) donors. In (C), CLL cells transfected as above were cultured for 48 h during which BrdU was added for the last 18 h of culture, or ³H-TdR was added for the last 24 h as detailed in Methods. Data from CLL-7 is shown.