Figure 4
The neutralizing effect of pro-Canakinumab on IL-1β downstream signaling. Luciferase activity of NF-κB promoter constructs (pNF-κB-Luc) transiently transfected into HEK293 cells with or without treatment with IL-1β (10 ng/mL). IL-1β neutralizing ability was analyzed by incubating cells with saline (gray), Canakinumab (orange), pro-Canakinumab (blue), MMP-9 pre-incubated Canakinumab (pink) or MMP-9 pre-incubated pro-Canakinumab (purple) and the luciferase activity was detected through a dual-luciferase reporter assay system (Promega, Madison, WI, USA) according to the manufacturer’s protocol. IL-1β treatment was used as a control for reporter activity. The results are expressed as fold changes of luciferase activity (relative light unit, RLU) over those of internal control Renilla–Luc reporter plasmid. All data are the mean ± SD of triplicate independent experiments (n = 3). Statistical significance was calculated by t test with NS, no significance; *P < 0.05.
