qRT-PCR to characterize a change in Runx1 expression in NPCs upon induction of differentiation.

Isolated NPCs were initially expanded and maintained in a culture medium containing EGF and bFGF (10 ng/ml each). Differentiation was induced by removing EGF from the culture medium, and changes in the expression levels of Runx1 (A) as well as Neurog2 (B) and Lmo3 (C), neuronal markers, were evaluated in the lysates collected at different time points after the removal of EGF. The primer pair of mRunx1_5F and mRunx1_6B was used for amplification of Runx1. See S1 Table for the specific sequences of each primer used in this study. (D) qRT-PCR to evaluate a change in Runx1 expression in NPCs upon treatment with TGF-β. Relative Runx1 mRNA levels in NPCs treated with 5 or 10 ng/ml TGF-β1 for 24 hours. (E) Relative Runx1 mRNA levels in NPCs treated with 10 ng/ml TGF-β1 for the indicated periods Error bars: standard deviation. n = 3~4/group. **: <0.01. Statistics: One-way ANOVA followed by Holm-Sidak’s multiple comparisons test.