Fig. 2: Sema7A restores the polarization and axon outgrowth in Satb2-deficient neurons also in vitro.
From: Semaphorin heterodimerization in cis regulates membrane targeting and neocortical wiring
a Examples of WT, Satb2 deficient, and Sema7A rescue conditions in E14 primary cortical neurons after 2 days in vitro (DIV2). b After 4 days in vitro (DIV4), axons can be identified in all conditions by their enrichment of TAU-1 and depletion of MAP2. Scale bar = 50 μm. c Box & whisker plots (whiskers: min, max, box spanning the interquartile range, and center line at median) of neuronal morphology at DIV2 (left column) and DIV 4 (right column). Data were non-normally distributed (D’Agostino & Pearson, Shapiro-Wilk tests). For all tests, Kruskal–Wallis test followed by Dunn’s Multiple comparison was used, where adjusted p values < 0.001 = ***, <0.01 = **, <0.05 = *. DIV2 longest neurite nneurons WT = 48, Satb2−/− = 54, Satb2−/− + Sema7A n = 37; DIV2 nneurons primary/end neurites WT = 40, Satb2−/− = 39, Satb2−/− + Sema7A = 37. DIV4 axon length nneurons WT = 20, Satb2 −/− = 39, Satb2−/− + Sema7A = 37. DIV4 nneurons primary/end neurites WT = 21, Satb2−/− = 39, Satb2−/− + Sema7A = 37. A full list of p values can be found in Supplementary Data S2. d Loss of Satb2 is associated with a random distribution of the centrosome which can be restored by Sema7A. (Top row): Representative images of DIV2 primary cortical neurons stained with the golgi marker Gm130 and centrosomal marker Cdk5rap2 (scale bar 25μm). Radar plots depict the circularized histograms of angle counts, where 360 degrees are binned in 30-degree increments. Measured blind, the polarity angle was measured using a standardized 100 × 100 pixel cross placed with the parallel axis centered in the process and the perpendicular axis touching the Golgi. Data were non-normally distributed (D’Agostino & Pearson, Shapiro–Wilk tests). Kruskal Wallis test followed by Dunn’s Multiple comparison padjusted GFP vs Cre <0.0001, padjusted GFP vs Cre + Sema7A = 0.0026, padjusted Cre vs Cre + Sema7A = 0.0794. nneurons WT = 35, Satb2−/− = 25, Satb2−/− + Sema7A = 35. For simplicity, panels denote p < 0.05 as *; p < 0.01 as **; and p < 0.001 as ***. All source data are provided in the Source Data file.
