Figure 6. Structure-based mutations disrupt MyD88^TIR:MyD88^TIR interactions.

a, Non-symmetrized projection map (from cryo-EM) of the MyD88^TIR assembly with a structure model of the MyD88^TIR assembly overlayed. b, Effect of MyD88^TIR alanine mutations on MAL-induced MyD88^TIR assembly. Turbidity was measured during lag phase (blue bars) and after 90 min (red bars) for the MyD88^TIR (60 µM) mutants after incubation with MAL^TIR (3.0 µM) at 30°C. Presented as mean ± range (N = 3). c, Effect of TIR-domain mutations on the ability of MyD88^FL to cluster in HEK293 cells. HEK293 cells were transfected with V5-tagged MyD88^FL (wild-type [WT] or mutant) and immuno-stained. Cells with a similar range of MyD88 expression levels were selected (lower panel). The percentage of cells with clustered MyD88 was quantified as in Fig. 1i and Supplementary Fig. 4d. The mean and range of duplicate transfections are shown (similar results were obtained in a second experiment).