Fig. 2. Regulation of nestin expression by rSCs by the serum. rSCs were cultivated in different culture medium: DEM/F12+N2+B27 (A), DEM/F12 (B), DEM/F12+N2 (C) or DEM/F12+B27 (D) and DEM/10% FBS (E). Cultures were labeled with anti-nestin antibody (green) and cell nuclei were counterstained with ethidium homodimer (red). Scale bar: (A-E) 40 µm. Nestin expression is present in cells grown in all serum-free media, and this result was confirmed by western blotting (F). Radial glial cells (RGCs) were used as nestin-positive control and ß-actin was used as protein-loading control.