Fig. 3: Structural and biochemical characterization of the WDR5–NANOG HD interaction.

a Crystal structure of the WDR5-NANOG HD complex. The complex was crystallized using the R100A mutant of NANOG HD and a truncated WDR5 (23-334). NANOG HD and WDR5 are shown in cyan and green, respectively. b Detailed interactions between the N-arm of NANOG HD and WDR5. Hydrogen bonds are indicated by yellow dashed lines, and key residues are shown as ball-and-stick models. A water molecule involved in the interaction is shown in red. c Detailed interactions between the C-terminal region of NANOG HD and WDR5. WDR5 is shown as an electrostatic surface, and interacting residues of NANOG HD are displayed as sticks. d Close-up view of the interaction between NANOG HD residue R153 and surrounding residues in WDR5. e NMR mapping of the WDR5-NANOG HD interface. Binding sites of NANOG HD and WDR5 are indicated in blue and red, respectively. f ITC binding curves for wild-type and mutant forms of NANOG HD and WDR5. Mutations at the binding interface markedly reduced or abolished the interaction. g Co-IP showing the interaction between full-length WDR5 and various full-length NANOG mutants. h. The kinetics of full length NANOG^WT, NANOG^WT + WDR5^WT, NANOG^WT + WDR5^F133A, and NANOG^W8A, measured using 5 μM NANOG and a 2:1 molar ratio of WDR5:NANOG; Buffer + ThT served as control. Traces show normalized ThT fluorescence over time. Data are mean ± SD from three independent experiments. Source data are provided as a Source Data file.