Figure 6

MyD88 TIR-domain validation of the target specificity of compound 12b. To determine whether SMA 12b inhibits MyD88-signaling by direct binding to the TIR domain, the SMA was pre-incubated at room temperature with varying amounts of TIR domain protein (or with BSA as control) for 2 h prior to being added to cells for testing for SEAP activity as described in Fig. 4. Data (a) are presented as mean ±SD SEAP response units from triplicate analysis of a single experiment where *p < 0.05, **p < 0.01 and ***p < 0.001 for conditions of interest versus LPS + 12b by 1-way ANOVA and Bonferroni post-test and (b) The log-log linear regression model is fitted to the data relating SEAP activity to TIR concentration as shown in equation 1 below by analysis carried out in SAS version 9.4 SAS proc MIXED. Equation: \(Log(\tfrac{SEAP\,Activity}{Activit\,of\,s5\,in\,DMSO})\) = \(\beta \,Log([TIR])+\alpha \).