Point mutation in NES2 induces nuclear accumulation of full-length Ngn3.

<p>(<b>A–F</b>) Cultured neurons were co-transfected with constructs encoding EGFP and full-length myc-tagged wild-type Ngn3 (myc-Ngn3), Ngn3 with leucine 135 mutated to alanine (myc-Ngn3-L135A) or empty vector expressing myc-tag. After 16 h, double immunostaining was performed using an anti-myc antibody to determine subcellular localization of wild-type and mutated myc-Ngn3 (A-C) and an anti-GFP antibody to visualize neurons at full (D-F). (<b>G</b>) Quantification of the relative fluorescence in the cell nucleus versus the cytoplasm of neurons. Graphs show the results (mean+s.e.m.) of at least three experiments. Significance levels were determined using a Student’s t-test; *** p<0.001 versus values of neurons transfected with plasmid expressing myc-tag.