Fig. 6
Actomyosin bias and vinculin tension were diminished in the Wnt5a−/− mutant mandibular arch. a Spatial distributions of F-actin and pMLC were not biased in 21 somite stage Wnt5a−/− mutants. The angular distribution of immunostain fluorescence intensity for epithelial (n = 4 embryos) and mesenchymal (n = 4 embryos) F-actin, and epithelial pMLC (n = 5 embryos) was quantified relative to the arch long axis that was designated 0o using SIESTA. Scale bars: whole arch panel 100 μm, merge field 20 μm, asterisks denote p < 0.05, Student’s t-test, error bars denote s.e.m. b Average fluorescence lifetime values in middle (mid) and distal (dist) epithelium and mesenchyme of mandibular arch. Boxplots show mean (x), median (---), central quartiles (coloured box), and range (end bars); n = 15 cells per region x 2 (mutant) or 3 (WT) embryos. p values for pairwise WT/Wnt5a−/− mutant comparisons are given in the graph. Of note, middle mesenchymal lifetime range was diminished in Wnt5a−/− mutants and more closely resembled WT distal mesenchymal lifetime values. c Representative vinculin force curves of an individual Wnt5a−/− mutant cell in the middle region that lacks the amplitude observed for WT cells in same region. d Multiple vinculin force curves; the variance (amplitude) of lifetime values for Wnt5a−/− mutant middle mesenchyme (0.0122 ns, n = 3 embryos, 15 cells per embryo) was diminished relative to WT middle mesenchyme (0.0201 ns, n = 5 embryos, 15 cells per embryo, p = 0.04), but similar to WT distal mesenchyme (0.0132 ns, p = 0.38, ANOVA). Lifetime variance was similar between WT (0.0132 ns) and Wnt5a−/− mutant (0.0096) distal mesenchyme (p = 0.13). e Variation of cytosolic calcium concentration using Fluo8 applied to embryos was quantified at 5 positions per cell over time. Corresponds to Supplementary Fig. 6D; scale bar 10 μm. Source data are provided as a Source Data file
