Figure 5.

Effect of human IRAK-M and death domain mutants in macrophages. WT IRAK-M and DD-mutants were stably expressed in the human monocytic cell line THP-1 by lentiviral transduction and FACS-sorting of EGFP positive cells which is bicistronically expressed. THP-1 cells were matured to macrophages before stimulation as described in Methods. (A) IRAK-M expression evaluated by western blotting. (B) The effect of stable WT IRAK-M expression and IRAK-M mutants was determined on TLR2 (Pam3CSK4) and TLR4 (LPS) mediated TNF and IL-6 production after stimulation for 6 hour. Shaded bars depict results of IRAK-M molecules with combinations of mutated residues/stretches within the same area. N=4, mean±SEM. *Indicates a difference with WT IRAK-M P<0.05.