Figure 3
Effects of bFGF and p21Cip/WAF1 siRNA on regulation of the VPA-induced differentiation and inhibition of proliferation in NPCs (A-B). NPCs were treated with 1 mM VPA for 48 h in the presence or absence of 10 ng/ml bFGF. (C-D) NPCs were transfected with 100 nM p21Cip/WAF1 siRNA prior to treatment with 1 mM VPA for 48 h in the presence of 10 ng/ml bFGF. Whole-cell lysates were subjected to immunoblotting to detect p21Cip/WAF1, Tuj1, or β-actin. Alternatively, cells were processed for immunofluorescent labeling to detect the presence of Tuj1 (green), p21Cip/WAF1 (green), or BrdU (red). Nuclei were counterstained with DAPI. Images are 400× magnifications. (E) NPCs were treated with 1 mM VPA for 48 h in the presence of 10 ng/ml bFGF. Cells were processed for immunofluorescent labeling to detect the presence of p21Cip/WAF1 (green) or BrdU (red). Nuclei were counterstained with DAPI. Magnification is 1200×.