Ifnγ signaling is up-regulated following constitutive TIRAP expression in murine HSPCs via a noncanonical pathway. (a and b) Gene set enrichment plots for Ifnγ response (a) and Il10 pathway (b) obtained from RNA-seq analysis of WT HSPCs transduced with TIRAP (n = 3) or vector (n = 3). NES, normalized enrichment score. (c) Validation of cytokine gene expression in vector- and TIRAP-transduced WT HSPCs by RT-qPCR. (d) Correlation between IFNγ gene expression and TIRAP gene expression in low-risk MDS patients; P = 0.0003. (e) Correlation between IL-10 gene expression and TIRAP gene expression in low-risk MDS patients; P = 0.12. Data were analyzed from a previously published microarray dataset (Pellagatti et al., 2006) for plots in panels d and e. (f and g) Structure of WT (f) and TRAF6-DN (g) RT-qPCR for Il10 and Ifnγ following TIRAP overexpression and knockdown of TRAF6 mutant. (h and i) Structure of WT (h) and MyD88-DN mutant (i) RT-qPCR for Il10 and Ifnγ following TIRAP overexpression and knockdown of MyD88. Data pooled from two to four independent experiments. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.