FIG. 2.
Depletion of Wee-1 by siRNA transfection leads to apoptosis. (A) Immunoblot of Wee-1. HeLa cells (1.5 × 105 cells per well of a six-well plate) were transfected with 21-nt siRNAs targeting Wee-1 or Lamin A/C as described in the text. Immunoblotting of cell lysates prepared at 24 h after transfection was performed with antibodies against Wee-1 or β-actin. Twenty micrograms of cell lysate was loaded in each lane and separated by SDS-10% PAGE, followed by immunoblotting with antibodies to Wee-1 or β-actin. (B) Cell proliferation after siRNA transfection. Cells were stained with 0.4% trypan blue and were counted under a microscope at different time points. (C) Apoptosis occurred following Wee-1 siRNA transfection. Apoptosis was measured by flow cytometric analysis of Annexin V-FITC- and 7-AAD-stained cells.