Extended Data Fig. 8: Stem Cell Pluripotency Assays.

a, Fluorescence microscopy images of overexpressed GFP-tagged NANOG WT (left) and W8A mutant in ESCs. Similar results were observed with 2 biological replicates. The characteristic stem cell colonies are maintained in WT but not in W8A mutant. b, Lower magnification fluorescence microscopy images of overexpressed GFP-tagged NANOG WT (left) and W8A mutant (right) in ESCs showing widespread differentiation in the mutant. Similar results were observed with 2 biological replicates. c, Crystal violet staining (stain all cells) of ESCs. Similar results were observed with 2 biological replicates. d, Alkaline Phosphatase (AP) staining of ESC colonies with overexpressed GFP-tagged NANOG WT (left) and W8A mutant (right). More AP+ colonies were observed with WT than mutant. Similar results were observed with 3 biological replicates.