Fig. 2

Altered tri-lineage differentiation of hESCs carrying H3K27M mutation. A Schematic of the neural induction scheme used (see the “Methods” section for details). B Representative bright field images of plated EBs at 3 and 5 days after initiating differentiation with neural induction media (scale bar=100μm). C Schematic of the tri-lineage differentiation scheme utilized for protein array analysis (see the “Methods” section). D Analysis of tri-lineage marker expression by protein array. Summary of fold differences between H3K27M and WT hESCs, when maintained in hESC/pluripotent stem cell (PSC) maintenance media or induced to undergo ectodermal, mesodermal, or endodermal differentiation. After normalization of intensity values on each array with internal controls, fold differences between the expression of each gene in the H3K27M versus the WT hESC line under each differentiation condition were calculated and averaged across arrays performed with six independent replicate sample sets utilizing two clonal lines. Clones used for each biological replicate experiment and numbers of biological replicate experiments performed are described in Additional file 2: Table S1