Figure 4.

Effect of PGA₁ on NF-κB- and HSF-dependent transcription. Jurkat cells were transfected with a CAT reporter gene construct controlled by the promoter/enhancer of HIV-1 (HIV-LTR wt) or a construct with mutations altering the two NF-κB binding sites in the LTR (HIV-LTR κB mu). Alternatively, cells were transfected with a CAT reporter gene construct containing the human HSP70 promoter sequence (LSN wt). After 24 h cells were divided into four aliquots and stimulated with 25 ng/ml TPA for 9 h in RPMI 1640 medium supplemented with 10% FCS in the presence or absence of 24 μM PGA₁: unstimulated (C) untreated (−) or PGA₁-treated (+); TPA-stimulated (TPA) untreated (−) or PGA₁-treated (+). The acetylated form of [¹⁴C]-1-deoxychloramphenicol (AC) was separated from unreacted reagent (CM) by ascending thin layer chromatography. An autoradiography is shown (Upper). Quantification of CAT activity is shown (Lower) and expressed as fold induction of untreated, unstimulated control. Results from three independent experiments are shown as average values.