NF-κB activation in human HEK293T cells transiently expressing TLR7 or TLR5 and the indicated human UNC93B1 alleles was measured using a dual luciferase reporter assay. Cells were stimulated with R848 (10 ng ml−1) or flagellin (2 ng ml−1) for 16 h before collection. Data are normalized to Renilla expression and expressed as RLUs. Data are mean ± s.d., n = 3 biological replicates. *P < 0.05, **P < 0.01 and ***P < 0.001, one-way ANOVA followed by a Tukey’s post-test (95% confidence interval). Data are representative of three independent experiments.