Figure 4.

SB203580 at 1 μM and kinase deficient MKK3 and MKK6 do not affect NF-κB transactivation potential. (A) TF-1 cells were transiently transfected as described in ‘Methods' with pIL6luc(-122) or pGAL4p65. Six hours after electroporation cells were stimulated for 24 h with medium, SB203580 (1 μM), OA (30 ng ml⁻¹), or SB203580 (1 μM, 30 min pretreatment) plus OA. Twenty-four hours after stimulation cells were lysed and analysed for the amount of produced luciferase protein. Promoter activity after treatment with medium is set at 1. Mean fold induction and standard error of the mean represent six identical experiments. ^*P<0.05. (B) TF-1 cells were transiently transfected as described in ‘Methods' with 15 μg pIL6luc(-122) together with 15 μg (1) pcDNA3 empty vector, (2) pRSV-MKK3(Ala), or (3) pcDNA3-MKK6(K82A) and 2 μg of a CMV-CAT construct. Six hours after electroporation cells were stimulated for 24 h with medium or OA (30 ng ml⁻¹). Twenty-four hours after stimulation cells were lysed and analysed for the amount of produced luciferase protein. The IL-6 promoter activity is represented in arbitrary luciferase units corrected for transfection efficiency. Mean and standard error of the mean represent at least three identical experiments.