Figure 4: A distinct balance of WNT and BMP signalling is required for cardiomyogenesis.

(a) Representative images showing the effect of additional medium refreshment after 6 h during the 24 h CHIR treatment on the NKX2.5-GFP expression measured on day 10 and (b) the analysis of six independent experiments thereof. (c) Heatmap showing induction of NKX2.5-GFP⁺ on day 10 following concomitant addition of indicated substances during the 24 h CHIR treatment. (d,e) Representative images on day 10 after concomitant addition of BMP4 to CHIR treatment and flow cytometric analysis thereof (n=3). (f) Representative flow cytometric data showing increase in MIXL1⁺ following concomitant BMP4 treatment after 24 h. (g,h) Effect of concomitant addition of either dorsomorphin (DM) (g) or IWR1 (h) to CHIR treatment on NKX2.5-GFP⁺ determined by flow cytometry on day 10 (n=4 of independent experiments in both graphs). All bars shown in this figure represent mean±s.e.m. *P<0.05, evaluated by one-way analysis of variance with post hoc Bonferroni analysis in all bar graphs. Scale bars, 1 mm. See also Supplementary Figs 5 and 6. NS, not significant.