Figure 3.

HNP-1 is required for NOD2-induced NF-κB pathway activation. (A) NF-κB luciferase assay was performed on HEK293 cells transfected with DNA encoding NOD2 (1ng), DNA encoding HNP-1 at different concentrations (10–100ng), NF-κB luciferase and Renilla reporter plasmids as well as empty vector. Cells were stimulated with 1 μg of MDP-LD for 18 h followed by measurement of luciferase and normalized with Renilla in cell lysates. *p<0.05 (B) Effect of specific siRNA against HNP-1 on ligand NOD2-dependent NF-κB activation. HEK293 cells were transiently transfected with the above mentioned plasmids and two different siRNA proven to knock down HNP-1 (100ng) as well as empty vector as control. *p<0.05 (C) HCT116 cells expressing endogenous NOD2 were transfected with two differents siRNA against HNP-1 and siRNA control and stimulated 36 h later with 1μg of MDP. NF-κB activation was measured 18 h after MDP stimulation as described above. *p<0.05