Fig. 1.

Establishment of EGFRviii-expressing human neural stem cells. a Phase-contrast images showing morphological characteristics of parental F3 cells and F3.EGFRviii cells after treatment with Doxycycline (DOX; 1 μg/ml) compared with mock (scale bar = 500 μm). b The mRNA expression of EGFRviii in F3 and F3.EGFRviii cells was determined by real-time PCR (n = 2). F3.EGFRviii cells were treated with DOX (1 μg/ml) at the indicated times. c F3 and F3.EGFRviii cells were treated with DOX (1 μg/ml) and harvested at the indicated times. The level of EGFR was determined by immunoblotting, using α-tubulin as a loading control. d F3.EGFRviii cells were immunostained with an antibody to pEGFR (green) after treatment with Dox (1 μg/ml). DAPI (4′,6′-diamidino-2-phenylindole; blue) was used for a nuclear counterstain. The white arrows indicate pEGFR-positive cells. e F3 and Dox-treated F3.EGFRviii cells (1 μg/ml) were harvested at the indicated times and subjected to immunoblot analysis using β-actin as a loading control