Fig 1: WNT11 treatment and analysis of WNT11 receptors in undifferentiated and differentiating ESCs. (A) Expression of Nanog, Pax6, Gata4 and T in WNT11-treated and control ESCs, EBs and EBOs cultured in medium supplemented with SR. Cells were collected after 48h of WNT11 treatment. (B) Expression of WNT11 receptors: Fzd5 and Fzd7. (C) FACS analysis: proportion of undifferentiated ESCs and cells obtained after disaggregation of EB2, EB5, and EB7 synthesizing FZD5 or FZD7. Data presented as means of three independent experiments with standard deviations. Representative histograms present proportion of EB5-derived cells synthesizing FZD5 or FZD7 or cells incubated with IgG only (negative control). Cells were cultured in medium supplemented either with SR or FCS. For qPCR analyzes (A,B) ß-actin was used as a reference gene. RQ = 1 for the level of gene expression detected in a 13.5-day-old mouse embryo. The average ?Cq for reference sample is shown on each graph. Data are presented as means of three independent experiments with standard deviations. * p < 0.05; Red color indicates differences between undifferentiated ESCs and EBs or EBOs; black color indicates differences between cells cultured in medium supplemented either with FCS or SR.