Fig. 3

JIP1 Phosphorylated at Thr205 enhances axonal outgrowth in CD-1 cultured cortical neurons and in vivo. A Representative images at DIV 3 of CD-1 cortical neurons infected with adenoviral GFP control (GFP), WT-JIP1 (JIP1), its alanine mutants (T205A and S235A), or its aspartic acid mutants (T205D and S235D). Scale bar, 50 μm. B Quantification of axonal length relative to GFP control was analyzed by one-way ANOVA and are represented as mean ± SEM; n=227 for GFP, n=191 for T205A, n=179 for T205D, n=156 for S235A, n=171 for S235D, and n=154 for JIP1. “n” equals the number of neurons from one experiment. Data representative of 3 independent experiments. C Representative images of axonal outgrowth in CD-1 embryonic brains subjected to in utero electroporation at E 14.5 with either the GFP control or T205A, T205D, or JIP1 plasmid. All constructs express GFP under a separate promotor. Brains were collected for axonal length analysis 2 days after electroporation. D Quantification of axonal length relative to GFP control analyzed by one-way ANOVA. Data presented as mean ± SEM. 30 to 55 neurons per mouse, 4 mice each for GFP and T205D and 3 mice each for T205A and JIP1, were used for quantification. Scale bar, 50 μm. * p < 0.05 and **** p < 0.0001