Figure 3
Inhibitors and dominant negative mutants targeting for NF-κB and AP-1 pathways attenuated LMP1-augmented human iE κ activities. (A) Bay11-7082 and SP600125 inhibited the activities of iEκ induced by stable expression LMP1. HNE2 and HNE2-LMP1 cells were transfected with pα-iEκwt, pGL3-α or pGL3-Basic vector, and pRL-SV40 as an internal control for transfection efficiency. 24hr after transfection, cells were treated with Bay11-7082 (20 μM), SP600125 (20 μM) or 0.1% DMSO for 12hr. Cells were harvested at 36 h after transfection and subjected to the luciferase assay. Statistical significance: * P < 0.01 and ** P < 0.05 vs. HNE2-LMP1 vehicle control. (B) Stable expression DNMIκBα and TAM67 inhibited the activities of iEκ increased by LMP1. Indicated NPC cell lines were transfected with pα-iEκwt, pGL3-α or pGL3-Basic vector, and pRL-SV40 as an internal control for transfection efficiency. Cells were harvested at 36 h after transfection and subjected to the luciferase assay. Statistical significance: * P < 0.01 and ** P < 0.05 vs. HNE2-LMP1 control. (C) Both DNMIκBα and TAM67 inhibited the activities of iEκ induced by transient expression LMP1. HNE2 cells were co-transfected with 400 ng/well of pα-iEκwt, pGL3-α or pGL3-Basic vector and 80 ng/well of internal control pRL-SV40 together with 200 ng/well of pSG5-LMP1, pDNMIκBα or pTAM67 expression plasmid. The total amount of DNA (~800 ng) was kept constant by addition of blank expression plasmid pSG5 necessary to normalize the amount of DNA transfected. Cells were harvested at 36 h after transfection and subjected to the luciferase assay. Statistical significance: # P < 0.01 vs. pSG5-transfected HNE2, * P < 0.01 and ** P < 0.05 vs. pSG5-LMP1-transfected HNE2.