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. 2019 Dec 9;219(1):e201902088. doi: 10.1083/jcb.201902088

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© 2019 Boyer et al.

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Figure 3. — TRIM67 is required for axon and growth cone responses to netrin-1. (A) Scanning electron micrographs of axonal growth cones from embryonic Trim67^+/+ and Trim67^−/− cortices treated with sham media or media containing netrin-1 for 40 min after 2 d in vitro. (B) Fluorescent micrographs of growth cones from primary neuronal cultures stained for filamentous actin (phalloidin) and β-III-tubulin. (C) Individual data points and box plots of growth cone responses after acute netrin-1 treatment (40 min), including increase in growth cone area, filopodial density, filopodia number, and filopodia length. Three experiments per genotype/treatment. n (cells) = 127 +/+ media, 103 +/+ netrin, 98 −/− media, 83 −/− netrin. For filopodia length, n (filopodia) = 729 +/+ media, 956 +/+ netrin, 786 −/− media, 1,076 −/− netrin. (D) Fluorescent micrographs of neurons cultured for 3 d in vitro including a final 24 h with addition of media or netrin-1, shown as the combined fluorescence of staining for both filamentous actin (phalloidin) and β-III-tubulin. (E and F) Individual data points and box plots of axon branching per 100-µm axon length (E) and of total axon length (F). Six experiments per genotype/treatment. n (cells) = 188 +/+ media, 186 +/+ netrin, 178 −/− media, 156 −/− netrin. *, P < 0.05; ***, P < 0.005; n.s., P > 0.05. Box plots are minimum, Q1, Q2, Q3, maximum.