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Fig. 8 | Molecular Medicine

Fig. 8

From: Lipopolysaccharide Signals Activation of Tumor Necrosis Factor Biosynthesis Through the Ras/Raf-1/MEK/MAPK Pathway

Fig. 8

Release of translational blockade imposed by the TNF 3′-UTR is prevented by dominant inhibitors of ras and raf-1.

RAW 264.7 cells were cotransfected with a reporter construct in which a CMV promoter was used to drive transcription of a CAT mRNA in which the TNF 3′-UTR was appended, together with empty vector (control) or vectors encoding dominant inhibitors of ras or raf-1. Cultures were induced with LPS at a concentration 1 µg/ml 24 hr after transfection, and cells were harvested for CAT assay 48 hr after transfection. Quantitation of CAT activity, and calculation of the induction ratio, was performed using a phosphorimager. CM, chloramphenicol; AcCM, acetylated forms of chloramphenicol.

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