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. 2005 Dec 5;171(5):857–869. doi: 10.1083/jcb.200507061

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Copyright © 2005, The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 7. — GAP-43 acts downstream of Ral. (A) A phosphomimetic GAP-43 protein, cGAP-43(S42D), restores branching in neurons plated on laminin and expressing dominant-negative Ral. Shown here are neurons expressing EGFP-F (top left), RalA28N (top middle), or RalB28N (bottom left) and RalA28N or RalB28N and cGAP-43(S42D) (right). Cells are stained with an anti-myc antibody to detect mutant Ral and an anti–cGAP-43 antibody. Bar, 50 μm. (B) Quantitative analysis of branching in cells plated on laminin and injected with single Ral isoforms or with Ral and cGAP-43(S42D) (means ± SEM: GFP, 3.47 ± 0.37; RalA28N, 1.74 ± 0.52; RalA28N + cGAP-43(S42D), 2.42 ± 0.31; RalB28N, 1.56 ± 0.41; RalB28N + cGAP-43(S42D), 3.72 ± 0.35; *, P < 0.04; **, P < 0.001). (C) SCG neurons were plated on polyornithine, microinjected, and left to express the indicated proteins for 5 h before laminin addition. Expression of dominant-negative Ral decreases branching compared with EGFP-F–expressing cells (top, compare middle and right with left). Branching is restored in cells coexpressing cGAP-43(S42D) (middle and bottom rows). Bar, 100 μm. (D) Quantitative analysis of branching in neurons initially plated on polyornithine and injected with single Ral isoforms or with Ral and cGAP-43(S42A) before laminin addition (means ± SEM: GFP, 2.64 ± 0.35; RalA28N, 1.05 ± 0.32; RalA28N + cGAP-43(S42D), 1.82 ± 0.40; RalB28N, 1.74 ± 0.30; RalB28N + cGAP-43(S42D), 2.73 ± 0.45; **, P < 0.01).