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Fig. 7
Enrichment of Vangl2 at anterior membranes of mediolaterally elongated cells. (A-C′) Live embryos at 90% epiboly stage (9hpf; A,A2) or 5s stage (11.7hpf; B-C′). mCherry- and GFP-Vangl2-expressing cells were transplanted from Tg(vangl2:GFP-Vangl2) embryos into the WT unlabeled host. Yellow arrowheads indicate the anterior cell membranes where Vangl2 is enriched. (D,E) GFP-Vangl2 and mCherry posterior/anterior membrane FI ratios of individual cells in the notochord at 90% epiboly stage (9hpf) and at 5-6s stage (11.7-12hpf) (D) or in the neuroectoderm at 5-6s stage (11.7-12hpf) (E). The blue bars represent the average posterior/anterior FI ratios for each condition. (F-I) Vangl2 asymmetry analysis using GFP-VANGL2 mosaic expression. (F) Modified image of 64-cell stage embryo showing the mosaic synthetic mRNA injection performed. (G) Live embryos mosaically expressing GFP-VANGL2 (green)/H2B-RFP (red) in notochord cells. (H) Plot Profile (Fiji) quantification of FI of two cells (white rectangle in G) at four different z-planes (confocal optical slices). (I) Comparison of GFP-VANGL2, GFP-Vangl2 and corresponding mCherry membrane FI ratios of individual cells in the notochord at 5-6s (11.7-12hpf) stage. *P<0.05, **P<0.01, ***P<0.001; ns, not significant; unpaired t-test with Welch′s correction. Scale bars: 20µm in A,A′; 10µm in B-C′.