Fig 1: MDP-dependent and -independent NF-?B trans-activation by NOD variants. A Primer information for C483W mutagenesis; B FLAG for NOD2 expression levels and ß-actin analyzed by western blotting are shown in the top column. HEK293T cells transfected with plasmids, containing 100 ng NF-?B reporter plasmid (pNF-?B-Luc), 30 ng expression construct of each human NOD2, 10 ng internal control for normalization of transfection efficiency (pRL-TK), and the corresponding mock vector, were cultured with or without 5 µg/mL MDP for 24 h. NF-?B activity were measured using Duo-Glo Luciferase kit (Promega, #E2920). R334W mutation of NOD2 and R311W SNP were used as positive and negative controls, respectively. Values represent the mean of normalized data (mock without MDP = 1) of triplicate cultures, and error bar indicated SD; and C A molecular model of NOD2 was generated and mapped with the position of C483W mutation
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