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. 2016 Nov 22;6:37681. doi: 10.1038/srep37681

Figure 6. ACTN4 played a pivotal role in migration induced by PO.

Figure 6

(A) Neutralizing Antibody Assays showed the role of ACTN4 playing in migration induced by PO. Neurospheres were plated in PLL or PO pre-coated 24-well plates and images were captured by phase contrast microscopy after 12 hours. Normal rabbit IgG was used as a negative control. (B) Summarized graph indicated the number of migration cells from neurospheres after neutralizing antibody assays, with PLL as control. ***P < 0.01, one-way ANOVA followed by Tukey’s post hoc test (n = 5 for each group). (C) Quantitative analysis of migration distance from neurospheres after neutralizing antibody assays, with PLL as control. ***P < 0.01, one-way ANOVA followed by Tukey’s post hoc test (n = 5 for each group). (D) Blotting bands showed downregulation of ACTN4 using siRNA transfection. Lipofectamine (Lipo) was used as a negative control. (E) The effect of ACTN4 on filopodia formation using phalloidin staining (green) and the cage-like microtubule structure with tubulin immunostaining (red) in neurospheres with/without ACTN4 downregulation using siRNA transfection, compared to PLL control. (F) Bar graph indicated the percent of filopodia formation. ***P < 0.01, one-way ANOVA followed by Tukey’s post hoc test (n = 5 for each group). (G) Quantitative analysis of average number of primary leading processes. ***P < 0.01, one-way ANOVA followed by Tukey’s post hoc test (n = 5 for each group). (H) Quantitative analysis of average number of secondary branches. ***P < 0.01, one-way ANOVA followed by Tukey’s post hoc test (n = 5 for each group).