Macrophage-Activating Lipopeptide-2 Requires Mal and PI3K for Efficient Induction of Heme Oxygenase-1
Figure 7
MALP-2 induces PI3K activation is regulated by the formation of a Btk, Mal, c-Src and p85α complex.
A. Cells were pretreated with or without LFM-A13 (100 µM), or PP1 (50 µM) for 1 h, and then incubated in the absence or presence of 5 ng/ml MALP-2 for 15 min. Cell fractions were prepared and subjected to Western blotting analysis with an anti-p-Akt antibody. Data shown represents three experiments (means ± SEM). *, P<0.05 and **, P<0.01 for significant difference between compared groups. B, C. Cells were transfected with siRNA of Mal or MyD88, and then treated with MALP-2 for 15, 60 or 120 min. Phosphorylated Akt was detected by Western blotting. *, P<0.05 for significant difference between compared groups. D. Cells were treated with MALP-2 for the indicated times, the cell lysates were subjected to immunoprecipitation using an anti-c-Src or anti-p85α antibody, and followed by immunoblotting with anti-Btk, anti-Mal, anti-MyD88, anti-c-Src or anti p85α antibody. All figures are representative of three independent experiments.