Figure 3.

The expression profile of TGF-β signalling pathway components and response genes in the EpiSCs. (a) Heat map showing different expression patterns of pathway components in undifferentiated (day 0) Mixl1-early, Mixl1-intermediate and Mixl1-late EpiSCs. (b)(i)–(iii) Heat maps showing the different response of (i) Mixl1-early, (ii) Mixl1-intermediate and (iii) Mixl1-late EpiSC-derived embryonic bodies cultured for 4 days in Activin A- or Nodal-supplemented medium, compared with the expression profile at day 0 of culture. (For gene lists of the heat maps, see electronic supplementary material, table S4). (c) Functional annotation of genes that are significantly upregulated in cells of Nodal or Activin A cultures based on clustered GO terms using DAVID. Examples of upregulated genes are listed separately for Nodal and Activin A treatment in Mixl1-early EpiSCs. (d) Heat map showing different expression patterns of response genes in undifferentiated (day 0) Mixl1-early, Mixl1-intermediate and Mixl1-late EpiSCs. (e)(i)–(iii) Heat maps showing the different expression patterns of downstream genes of (i) Mixl1-early, (ii) Mixl1-intermediate and (iii) Mixl1-late EpiSC-derived embryonic bodies cultured for 4 days in Activin A- or Nodal-supplemented medium, compared with the expression profile at day 0 of culture. (For gene lists of the heat maps, see electronic supplementary material, table S4). (f) Functional annotation of genes that are differentially upregulated in Mixl1-early EpiSCs of Nodal or Activin A cultures. Examples of upregulated genes are listed separately for Nodal and Activin A treatment. The order that genes are listed on the heat map is determined by the clustering of the expression data, which were normalized against that of housekeeping genes: Actb, B2M, Gapdh, Hsp90ab1 and Gusb. Mixl1-E, Mixl1-early; Mixl1-I, Mixl1-intermediate; Mixl1-L, Mixl1-late; EB0, day-0 embryoid bodies; EB4, day-4 embryoid bodies; ctrl, control; ActA, Activin A. (Online version in colour.)