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. 2014 Apr 28;205(2):217–232. doi: 10.1083/jcb.201311003

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© 2014 Winkle et al.

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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — TRIM9 is required for Netrin-1–dependent branching. (A) E15.5 TRIM9^+/+ and TRIM9^−/− cortical neurons were untreated or Netrin-1 stimulated and stained with fluorescent phalloidin. Arrowheads denote branch points. (B) Mean axon branch density ± SEM. n > 80 neurons/condition. Expression of Myc-TRIM9 (T9) in TRIM9^−/− neurons reduced constitutive branching and rescued Netrin-1 sensitivity. MycTRIM9ΔRING (T9ΔRING) or MycTRIM9LD (T9LD) introduction failed to rescue Netrin-1–dependent branching. (C) E15.5 TRIM9^−/− cortical neurons expressing either Myc or MycTRIM9ΔRING were untreated or FGF2 stimulated and stained with fluorescent phalloidin and antibodies to Myc and βIII-tubulin (not depicted). Arrowheads denote branch points. (D) Mean axon branch density ± SEM. Stimulation of TRIM9^−/− neurons with FGF2 fails to increase already exaggerated axon branching except in cells expressing MycTRIM9ΔRING. (E) Axon length is not affected by Netrin-1 or genetic loss of TRIM9. Means ± SEM. All graphs are n > 80 neurons/condition from more than three independent experiments. P-values were obtained from ANOVA with Tukey’s post-hoc analysis.