Fig. 3.

Wnt/β-catenin activity is important for efficient self-renewal of naïve hESCs. (A–C) Graphs of the relative number of naïve (A and B) or primed (C) ELF1 hESCs grown on Matrigel with the indicated small molecules. Cells were allowed to attach for 16–20 h before the addition of DMSO, XAV, or IWP2. The media was changed daily. Cell number was measured using the CyQUANT assay at specified times. Fold changes were calculated relative to the cell number at the time the small molecules were first added (defined as 0 h). (D) Graph of the relative number of naïve or primed ELF1 hESCs transfected with the indicated siRNAs and grown on Matrigel for 3 d. Cell number was measured using CyQUANT assay and normalized to the value for Ctrl siRNA-transfected cells. (E) Graph of the relative colony formation potential of naïve ELF1 hESCs. Cells were cultured on Matrigel and with the indicated small molecules for 3 d, then lifted and plated onto MEFs at clonal density (250 cells per cm²), and cultured with the indicated small molecules for 3 more days. Colonies were subsequently stained for AP activity, and the number of AP-positive colonies was manually counted. The percentage is calculated as the number of colonies divided by the number of cells seeded (all of the data are means, and error bars represent S.E.M of n = 3 biological replicates; *P < 0.05 by t test; n.s., not significant).