Wnt/β-catenin signalling stabilises and enhances spontaneous symmetry-breaking and polarisation events in gastruloids. Wnt/β-catenin signalling stabilises.

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Wnt/β-catenin signalling stabilises and enhances spontaneous symmetry-breaking and polarisation events in gastruloids. Wnt/β-catenin signalling stabilises and enhances spontaneous symmetry-breaking and polarisation events in gastruloids. (A) T/Bra::GFP expression in gastruloids at 24 and 48 h prior to the Chi pulse (left), and examples of gastruloids following a DMSO or Chi pulse (n=28). Chi-mediated stimulation increases the robustness of the response and reproducibility of the phenotype. (B) Quantification of T/Bra::GFP reporter expression in individual gastruloids over time following DMSO (n=28), Chi (n=28) or Wnt3a (n=14) treatment. The maximum length of each gastruloid is rescaled to 1 unit and the fluorescence is normalised to the maximum fluorescence from the Chi treatment. The Wnt3a condition is from a different replicate (indicated by dashed horizontal line). Vertical line in each plot marks the peak max and the corresponding coordinates denote the position of this value. (C) Statistical analysis of the indicated treatments showing the normalised root square distance as a measure of the heterogeneity for each condition within each time-point, and the indicated P values as assessed by non-paired Student's t-test. Red line indicates the median, the 25th and 75th percentiles are denoted by the bottom and top edges of the box, the whiskers extend to the most extreme data points, and outliers are indicated by the plus symbol. (D) Heat maps indicating the average fluorescence (fluorescence norm.), the average area taken up by the standard deviation (StDev Area), average length and the roundness of the gastruloids after the indicated conditions and time-points from the traces in B (Fig. S2 and Materials and Methods). (E) Live imaging of a gastruloid subjected to a pulse of DMSO (top) or Chi (bottom) between 48 and 72 h AA (n=21/condition). Gastruloid length is indicated by the y axis (posterior=0 μm), time on the x axis and the fluorescence intensity in colour. Early time-points (24-72 h AA) were imaged using a higher power objective. Scale bars: 50 μm (pre-pulse); 100 μm (post-pulse). David A. Turner et al. Development 2017;144:3894-3906 © 2017. Published by The Company of Biologists Ltd