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. 2014 Apr 30;289(27):18987–18998. doi: 10.1074/jbc.M114.557686

FIGURE 3.

FIGURE 3.

HSP70 regulates NOD2 induced NF-κB signaling. A, dual-luciferase assay performed on HEK293T/NOD2Myc and HCT 116 transfected with HSP70 or control vector in the presence or absence of 2 μm MDP or 200 nm of TNFα incubated for 6 h. 75 ng of pGL432 and 7.5 ng of pRL vector was used per 24-well dish for transfection. Relative luciferase activity of firefly to Renilla is plotted. Results shown are the means ± S.D. of experiments performed in quadruplicate. *, p < 0.05 was considered as significant. B, dual-luciferase assay performed on HEK293T/NOD2Myc and HCT 116 cells incubated with 100 μm KNK437 or DMSO in the presence or absence of 2 μm MDP or 200 nm TNFα. 75 ng of pGL432, 7.5 ng of pRL vector, and 200 ng of HSP70 or control vector were used per 24-well dish for transfection. Relative luciferase activity of firefly to Renilla is plotted. Results shown are the means ± S.D. of experiments performed in triplicate. *, p < 0.05 was considered as significant. C, quantification using ELISA of the amount of IL-8 secreted by HCT 116, THP-1, and HEK293T/NOD2Myc cells incubated with 2 μm MDP or 200 nm TNFα in the presence of 100 μm KNK437 or DMSO. Results shown are the means ± S.D. of experiments performed in triplicate. *, p < 0.05 was considered as significant. D, quantification using ELISA of the amount of IL-8 secreted by HEK293T/NOD2Myc and HCT 116 cells transfected with HSP70 or control vector incubated with 2 μm MDP or 200 nm TNFα. Results shown are the means ± S.D. of experiments performed in triplicate. *, p < 0.05 was considered as significant.