Figure 3

Knockdown of endogenous Npas4 protein during neural differentiation of mESCs. (A) Immunoblotting was used to determine the level of Npas4 protein expression in different mESC lines (n = 3 biological replicates). Lysates were harvested after four days of differentiation in N2B27 medium and blots were probed with an anti-Npas4 antibody. β-actin served as a loading control. (B) Npas4 knockdown was quantified by densitometry analysis. Npas4 protein expression in each sample was normalized to β-actin expression and is relative to Npas4 expression in the Empty vector control line which was given an arbitrary value of 1. Mean values and standard deviations of three independent experiments (n = 3 biological replicates) are shown (**P <0.01, one-way ANOVA). ANOVA, analysis of variance; mESC, mouse embryonic stem cells.