Fig. 4

MyD88(114-118)Ala (Ala2) mimics the dominant negative effect of MyD88S. A Effect of overexpression of indicated MyD88 variants on IL-1β (10 ng/ml) and TNFα (100 ng/ml) induced stimulation of NF-κB responsive reporter expression tested in the presence of endogenous MyD88 in HEK293T cells. Data are normalized to the response of empty vector transfected and cytokine stimulated cells, and are shown as means ± SD of triplicate samples. Statistical significance was determined by Student’s t-test (*p < 0.05, **p < 0.001). B Effect of overexpression of indicated Flag-tagged MyD88 variants on the phosphorylation state of selected MyD88 effectors in HEK293-I3A cells. C Interaction of MyD88 variants (Flag-tagged) with endogenous IRAK4 was analyzed by co-immunoprecipitation on anti-Flag beads and evaluated using anti-Flag and anti-IRAK4 antibodies. D Diagram of simplified MyD88 signaling