Figure 1

Nlg1 clusters were mobile in dendrites of cortical neurons. (A) Image of a live neuron transfected with GFP-Nlg1 (green) and surface-labeled with anti-GFP Fab fragment (red). The arrowhead denotes one of the few clusters of GFP-Nlg1 that was not labeled by the Fab fragment. Scale bar, 5 μm. (B) Plot of the fluorescence intensities (in arbitrary units (a.u.)) for GFP-Nlg1 and anti-GFP Fab fragment from five neurons, showing strong correlation of intensities. (C) Time-lapse imaging of 4 d.i.v. neurons transfected with CFP-Nlg1 revealed Nlg1 clusters that were mobile in the dendrites (pink and black arrowheads) and filopodia (arrow). Immobile clusters are indicated by asterisks in the first panel. Time is in minutes and seconds. Scale bar, 10 μm. (D) Diagram depicting the principle interaction domains of Nlg1 and the deletions of the carboxy-terminal 4 and 55 amino acids (ΔC4, ΔC55). AChE, acetylcholinesterase domain; PDZb, PDZ binding motif; WW, ww interaction domain; Wt, wild type. (E) Images of live transfected neurons demonstrated the punctate distribution of GFP-Nlg1 (wild type (wt)) and GFP-Nlg1ΔC4 (ΔC4). The localization of GFP-Nlg1ΔC55 (ΔC55) was diffuse and showed very few clusters.