Transfection with siRNA reduces targeted protein over time.

<p>HeLa cells were untransfected (Un) or transfected with 25 nM siRNA to the indicated gene. Protein samples were subjected to Western immunoblotting and membranes probed for the protein of interest. Membranes were probed for Ran-GTP (Ran) as a loading control. Size markers in kDa are indicated to the left of each blot. At 24, 48, 72 and 96 hours post transfection (hpt), total protein was collected (A) or cells were fractionated into cytoplasmic (cyt) and nuclear (nuc) fractions (B). Panel C, HeLa cells were transfected with 0–25 nM siRNA to Wee1 and total protein collected at 24 hpt.