Figure 5: Sema3B invalidation alters the orientation of neural progenitor divisions at E10.5. | Nature Communications

Figure 5: Sema3B invalidation alters the orientation of neural progenitor divisions at E10.5.

From: Cerebrospinal fluid-derived Semaphorin3B orients neuroepithelial cell divisions in the apicobasal axis

Figure 5

(a) Cumulative fraction curves showing the distribution of neural progenitor division orientation in E10.5 Sema3B+/+ and −/− spinal cords (n, number of cells from a minimum of five embryos per condition, Sema3B+/+ and −/− embryos from the same litters were analysed; P value is from the KS test). Previous data sets are also displayed in pie charts (P value is from Fisher’s exact test). (b) Confocal images of representative anaphase cells with hoechst, PH3 and γ-tubulin stainings. (c) Planar and oblique division proportions in the ventral and dorsal halves of Sema3B+/+ and Sema3B−/− spinal cords. (d) PH3 immunolabelling revealing mitotic cells in E10.5 Sema3B+/+ and −/− spinal cord sections. (e) Mitotic index in E10.5 Sema3B+/+ and −/− spinal cord sections at the thoracic and brachial levels (n, number of sections from 10 embryos; ns not significant with Mann–Whitney U-test). The number of mitotic progenitors is not altered by Sema3B invalidation at E10.5. (f) In situ hybridization of Tis21 mRNA in E10.5 Sema3B+/+ and −/− spinal cord sections. (g) Tis21 mRNA staining area and intensity at the thoracic and brachial levels (n, number of cross-sections from four embryos in each condition; P value is from Mann–Whitney U-test). (h) Proportions of prophases (Pro), metaphases (Meta) and anaphases (Ana) among mitotic progenitors of E10.5 Sema3B+/+ and −/− spinal cords at the thoracic and brachial levels. The percentage of metaphase cells is increased in E10.5 Sema3B mutants (n, number of PH3+ cells analysed from five embryos for each conditions. Sema3B+/+ and −/− embryos were from the same litters). (i) Confocal images revealing ZO1 (planar view), aPKC (cross-section) stainings at the level of neural progenitors apical feet and F-actin and acetylated tubulin stainings in anaphase and metaphase neural progenitors (planar view) in Sema3B+/+ and −/− E10.5 spinal cords. Apical foot morphology, cortical actin and mitotic spindles are detected with similar patterns under both conditions. Scale bars in b,i, 5 μm, in d,f, 100 μm.

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