Fig. 4 | Nature Communications

Fig. 4

From: High-throughput three-dimensional chemotactic assays reveal steepness-dependent complexity in neuronal sensation to molecular gradients

Fig. 4

Neuronal response to Sema3A gradients with varied steepness. a Side and top view of 3D cultured neurons (β-tubulin) in response to Sema3A gradient of varied steepness, scale bar, 100 µm. b Quantitative analysis of neuronal migration and related association with the Sema3A gradient steepness, n = 6, error bars indicate standard deviation. c Box-plots for quantitative analysis of neurite repellence in response to varied Sema3A gradient steepness. More than 40 neurites (as indicated on top of each box) were pooled from four biological replicates. d Representative views of axonal (Tau-1+) or dendritic (MAP2+) differentiation under Sema3A gradient of decreasing steepness. The combined images of the indexed subpanels were enlarged and shown in the boxed region. Scale bar, 100 µm. e, f Box-plots for quantitative analysis of dendritic (e) or axonal (f) guidance in response to varied Sema3A gradient steepness. More than 20 neurites (as indicated on top of each box) were pooled from four biological replicates. For c, e, f, the parts of the box indicate 25, 50 and 75 percentiles, and the whiskers indicate 5% and 95%. The square mark indicates mean of the data. For b, c, e and f, the red line indicates a logarithmic fitting of the data mean; neuronal growth pattern in each hydrogel cylinder was compared in pairwise to experiments without any chemotactic factor treatment (Blank-ctrl, Supplementary Fig. 6), * or ** indicates a p-value < 0.05 or <0.005 by paired Kruskal–Wallis tests

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