FIG. 2.
Effect of novobiocin on the expression of TNF-α mRNA. Monocytes (2 × 106/ml) were incubated for 10 min with various concentrations of novobiocin or nicotinamide before 100 ng of LPS per ml was added for 2 h. mRNA was isolated and reverse transcription-PCR was then performed with primers specific for TNF-α and β-actin. Data represent the results of one of four representative experiments. Lanes: 1, unstimulated monocytes; 2, LPS-stimulated monocytes; 3 to 5, LPS-stimulated monocytes in the presence of 0.1, 0.25, and 0.5 mM novobiocin, respectively; 6, LPS-stimulated monocytes in the presence of 25 mM nicotinamide. The controls were no cDNA (lane −) and defined cDNA of the concerned protein (lane +). Lane M, marker (DNA molecular weight marker VI).