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. 2020 Feb 19;39(6):e102513. doi: 10.15252/embj.2019102513

Figure EV3. The fast transport of pre‐miR‐181a‐1 is decreased upon Sema3A exposure.

Figure EV3

  • A
    Schematic representation of the experimental protocol. Two hundred to two hundred fifty nanogram per microliter cy3‐pre‐miRNA‐181a‐1, 100 ng/ml Netrin‐1, 200 ng/ml Sema3A, and 200 ng/ml Slit2 or no cue control (1× PBS) were bath applied to the cultures for up to 7 min. Zero minute in the graph corresponds to time‐lapse movies acquired before treatment. The same axon was analyzed for each condition at 0, 3, 5, and 7 min.
  • B–D
    Frequency distribution of tracked puncta. Each data point corresponds to one axon. Total number of puncta analyzed is as follows: 244, 221, 211, and 158 (no cue at 0, 3, 5, and 7 min); 231, 245, 203, and 178 (Slit2 at 0, 3, 5, and 7 min); 133, 160, 119, and 85 (Netrin‐1 at 0, 3, 5, and 7 min); 125, 134, 106, and 111 (Sema at 0, 3, 5, and 7 min). Total number of axons analyzed is as follows: 11 (no cue), 9 axons (Slit2), 7 axons (Netrin‐1), and 7 axons (Sema3A). n = 6 (no cue), n = 3 (Slit, Netrin), and n = 4 (Sema) independent experiments. Values are mean ± SEM. ns, not significant.
  • E
    Average velocity of puncta. Each data point corresponds to one axon. Total number of puncta and axons analyzed is as follows: 150 puncta and 11 axons (no cue); 99 puncta and 7 axons (Sema3A). n = 4 (Sema3A) and n = 6 (no cue) independent experiments. Values are mean ± SEM. ns, not significant.
  • F
    Frequency distribution of tracked puncta. Each data point corresponds to one axon. Total number of puncta and axons analyzed is as follows: 150 puncta and 11 axons (no cue); 99 puncta and 7 axons (Sema). n = 4 (Sema) and n = 6 (no cue) independent experiments. Values are mean ± SEM. ns, not significant.
Data information: *P < 0.05, **P < 0.01. Data were normally distributed (B–D, F). Two‐way RM ANOVA (B). Two‐way ANOVA followed by Sidak's multiple comparison post hoc test (C, D). Data were not normally distributed. Kruskal–Wallis followed by Dunn's multiple comparison post hoc test (E). Two‐way ANOVA followed by Tukey's multiple comparison post hoc test (F).