UBXN1 inhibits TNFα-triggered NF-κB signaling at TNF receptor level.
A and B, UBXN1 inhibits the process of TNFα-triggered NF-κB activation. A, U2OS cells (2 × 105) were transfected with control siRNA, UBXN1-#1, and UBXN1-#2. After 72 h, the cells were treated with TNFα (10 ng/ml) for indicated times. The whole cell lysate was analyzed by immunoblotting with the indicated antibodies. B, HEK293T cells (2 × 105) were transfected with either an empty vector or FLAG-UBXN1 plasmid. After 36 h, cells were treated with TNFα (10 ng/ml) for the indicated times. The whole cell lysate was analyzed by immunoblotting with the indicated antibodies. C, UBXN1 inhibits TRADD-, TRAF2-, and RIP1-mediated but not TAK1- or IKKβ-mediated NF-κB activation. HEK293T cells (1 × 105) were transfected with reporter plasmid, either an empty vector, or FLAG-UBXN1 plasmid together with TRADD, TRAF2, RIP1, TAK1, IKKβ, respectively. Luciferase assay was assessed 24 h after transfection.