FIG 6.

Effect of GPS1 downregulation on NF-κB signaling. (A) The activity of the NF-κB signaling pathway was assessed by using a luciferase assay. HEK293 cells were treated with AllStars siRNA or siRNA for GPS1. Twenty-four hours later, an NF-κB reporter plasmid encoding luciferase and an NF-κB signaling pathway-activating protein (i.e., MEKK)-expressing plasmid were transfected; luciferase activities were measured 24 h after the plasmid transfection. The tests were performed in triplicate, and the error bars represent the standard deviation for triplicate samples. The statistical analysis was carried out by using ANOVA, followed by Sidak's multiple-comparison test. ***, P < 0.001 for three independent experiments. (B) Recovery of NF-κB signaling pathway activity in GPS1-downregulated cells. Twenty-four hours after siRNA treatment of HEK293 cells, an siRNA-resistant GPS1-expressing plasmid and an NF-κB reporter plasmid encoding luciferase were transfected into the cells. Forty-eight hours later, luciferase activity was measured. The luciferase activity of the AllStars siRNA-transfected cells was set at 100%. The tests were performed in triplicate, and the error bars represent the standard deviation for triplicate samples. Statistical analysis was carried out by using ANOVA, followed by Sidak's multiple-comparison test. **, P = 0.002 for three independent experiments.