Macrophage-Activating Lipopeptide-2 Requires Mal and PI3K for Efficient Induction of Heme Oxygenase-1
Figure 1
HO-1 expression in response to MALP-2 is dependent on TLR2 and TLR6.
A. THP-1 cells were preincubated with anti-TLR2 (10 µg/ml) or anti-TLR6 (1 µg/ml) for 1 h, and then stimulated with MALP-2 (5 ng/ml) for 16 h. HO-1 protein expression was analyzed by Western blot, and then densitometric analysis was performed after normalization with β-actin protein levels. B. Cells were transiently transfected with a dominant negative (DN)-TLR2 or DN-TLR6. After 20 h of transfection, cells were incubated with 5 ng/ml of MALP-2 for another 16 h. The cell lysates were prepared and separated by SDS-PAGE for Western blot analysis of HO-1. C. Cells were cotransfected with a DN-TLR2 or DN-TLR6 and HO-1-luc reporter gene, and then stimulated with MALP-2 (5 ng/ml) for 8 h. The luciferase activity derived from HO-1 activation was normalized the transfection efficiency with β-gal, and pZERO was used as a control vector. Data shown represent results from three independent experiments (means ± SEM). *, P<0.05 and **, P<0.01 for significant difference between compared groups.