Fig. 1

Human embryonic stem cells with H3K27M mutation have increased clonogenic stem cell properties. A Percentages of stem cells with clonogenic potential were assessed by plating on low attachment plates to promote sphere formation at extreme limiting dilution (ELDA). Data shown is an average +/− sem of four biological replicates utilizing four H3K27M clonal lines, each performed in technical triplicate. B hESCs were grown as 3-D spheres in maintenance media for 72 h and cell number was assessed by using the CellTiter-Glo assay (Promega, see Methods). Clones used and biological replicates performed are in Additional file 2: Table S1, with each biological replicate performed in technical triplicate, and WT values set at 1.0 for comparison with the K27M sample. C Representative bright field image of WT and H3K27M hESCs after 72 h of growth in non-adherent culture in maintenance media (scale bar=100μm). D 120 hESCs were plated under non-adherent conditions in maintenance media and the number of spheres that formed was assessed at day 7. Clones used for each biological replicate experiment and numbers of biological replicate experiments performed are described in Additional file 2: Table S1 for each finding in this manuscript. p values were calculated using an unpaired t-test