Figure 5

Knockdown of KLF2 modulates the imatinib induced apoptosis. A: HMC-1 cells were transfected with control siRNA or siRNA directed against KLF2 (KLF2 siRNA). 48 h after transfection, cDNA of untransfected (-) and transfected cells (control siRNA and KLF2 siRNA) was prepared and analyzed for expression of KLF2 and actin (ACTB) by semi-quantitative RT-PCR. KLF2 bands were quantified using TINA2.0 software and were normalized to actin. B: SiRNA transfected cells as in A were treated with imatinib (5 μM) for the indicated times. Every 3 h, whole cell lysates were prepared for detection of cleaved caspase-3 (cl. Caspase 3) and GAPDH by immunoblot. C: Cells were transfected and treated with imatinib as in B. At the indicated times after imatinib treatment, cells were fixed with paraformaldehyde and apoptotic cells were detected by TUNEL staining. Observations from different fields in the microscope (between 300 and 700 cells per experimental condition) are depicted as the mean +/- SD of % TUNEL positive cells. Student's t-test was applied to describe the significance of differences between cells treated with control siRNA and KLF2 siRNA. P-values ≤ 0.05 were considered as significant and are indicated above the corresponding columns. bp: base pairs of the DNA marker. MW: molecular weight.