Figure 6

Relative expression of IL-8, IL-6, ICAM-1 and IκBα genes in HeLa cells expressing IE63, IE63-S224/T222A, and IE63-Full versus control cells stimulated or not with TNFα. HeLa cells expressing IE63 wild-type, IE63-S224/T222A, IE63-Full or IE63 in inverted orientation (Inv, control) were treated for increasing times (from 0 to 2 h) with TNFα at a final concentration of 200 U/mL. (A) Total RNA extracts were isolated and analyzed by Real-time RT-PCR using primers for the IL-8 mRNA, IL-6 mRNA, ICAM-1 mRNA, and IκBα mRNA. (B) The IκBα degradation. HeLa cells expressing IE63wt or control cells were treated for increasing times (from 0 to 2 h) with TNFα (200 U/mL). IκBα degradation was followed by Western Blotting on total cellular extracts. β-actin Western Blotting detection was used as loading control (lower panel). ρ-values were calculated using the graphpad quickcalcs software [59]: *, significantly different from control (Inv ; p-value < 0.05) ; **, not significantly different from control (Inv ; p-value ≥ 0.05).