Figure 3
From: Enterovirus 71 2C Protein Inhibits NF-κB Activation by Binding to RelA(p65)
Mapping the region in 2C interacted with p65 IPT.
(A) The diagram of 2C truncated constructs. Numbers indicated amino acid position. (B) IPT domain of p65 interacts with 2C 1-125aa and 126-263aa. IPT-GST immobilized on glutathione-Sepharose beads were incubated with lysates from 293T cells transfected with 2C truncated constructs. The bound proteins were subjected to Western blots using indicated antibodies. (C) 2C truncated forms inhibit NF-κB activation. 293T cells were transfected with pNF-κB, pRL-TK and 2C truncated constructs for 24 hours and then treated with TNF (10 ng/ml) for 6 hours. The cells were assayed for dual luciferase activity. Asterisks indicate significant differences between groups, data statistics were used student t-test (mean ± SD, *** indicated p < 0.001). (D) 2C 122-125aa is required for binding to p65 IPT domain. IPT-GST immobilized on glutathione-Sepharose beads were incubated with lysates from 293T cell transfected with 2C truncated constructs. The bound proteins were subjected to Western blots using indicated antibodies.
