Fig. 4.

Transcriptional regulatory potentials of TRAFs 1–7. a NF-kB dual-luciferase assay performed on Neuro2a (light gray) and HEK 293 (dark gray) cell extracts , with the overexpression the pBind vector, pBind-TRAF2, or pBind-ΔN-TRAF2. Neuro2a and HEK 293 cells were transfected either with the control pBind vector or with pBind-TRAF2 or pBind-ΔN-TRAF2 (1 μg each). After 24 h, the cells were lysed and dual-luciferase assays were performed. b 5x Gal4 promoter-driven dual-luciferase assay performed on Neuro2a and HEK 293 cell extracts, with the overexpression of the pBind vector, pBind-TRAF2, or pBind-ΔN-TRAF2. c 4x Gal4-minimal TK promoter-driven dual-luciferase assay performed on Neuro2a and HEK 293 cell extracts, with the overexpression of the pBind vector or pBind-TRAF1-7. d 4x Gal4-minimal TK promoter-driven dual-luciferase assay performed on Neuro2a and HEK 293 cell extracts, with the overexpression of the pBind vector, pBind-TRAF2, or pBind-ΔN-TRAF2. e 5x Gal4 promoter-driven dual-luciferase assay performed on Neuro2a and HEK 293 extracts, with the overexpression of the pBind vector or pBind-RelA, in turn with either the pCMV-Tag2C vector or pCMV-Tag2B-TRAF1-7. f 5x Gal4 promoter-driven dual-luciferase assay performed on Neuro2a and HEK 293 cell extracts overexpressed with pBind RelA, in turn coexpressed with the pCMV-Tag2B vector, pCMV-Tag2B-TRAF2 or pCMV-Tag2B-ΔN-TRAF2. Each of these experiments was repeated at least three times, and the data represent the mean of at least four independent sample readings from one representative experiment ± the SEM, normalized to the pBind Renilla luciferase control readings. One-way ANOVA was performed, followed by Tukey’s test; *≤0.05; **≤0.001. VC vector control, T1 TRAF1, T2 TRAF2, T3 TRAF3, T4 TRAF4, T5 TRAF5, T6 TRAF6, T7 TRAF7