Figure 5

Co-expression of AmphiSoxE and Snail2 causes disruption of apical-basal polarity of neuroepithelial cells.

(a,b,e,f) Transfection of AmphiSoxE or Sox9 alone: Apical N-CAD expression is reduced as indicated by white arrows. (c,d,g,h) Most of the transfected cells are retained in the neural tube except for the dorsal and medial regions where basal LAMININ expression is reduced (white arrows). (i,j,k,l) Cotransfection of AmphiSoxE and Snail2 assayed at 24 hours post-transfection. (i,j) The apical N-CADHERIN expression is lost (white arrow). (k,l) Many cells have delaminated from the neural tube and basal LAMININ expression is lost (white arrow). (m–p) A similar observation is seen in the neural tube transfected with Sox9 and Snail2, with loss of N-CAD and LAMININ expression (white arrows). Scale bar: 100 μm. Transverse section of the neural tube electroporated with EGFP (q), AmphiSoxE/EGFP (r) and Sox9/EGFP analyzed at 48 hours post-transfection. Early onset of migration onto the dorsolateral migratory pathway is observed in AmphiSoxE/EGFP⁺ (r) or Sox9/EGFP⁺ cells(s) as indicated by the white arrows, but not observed for control EGFP⁺ cells (q). These dorsolateral migrating cells do not express the melanocyte marker, MelEM (r,s). Insets show a low magnified view of panels q-s with yellow boxes indicating the high magnification of the region where MelEM immunofluorescence is shown in each panel. (t) Quantification of the number of EGFP⁺, AmphiSoxE/EGFP⁺ and Sox9/EGFP⁺ cells migrating via the dorsolateral route. nt, neural tube. ***p < 0.001 as compared to EGFP control. Scale bar: 20 μm.