FIGURE 3.

PE_PGRS11 triggers TLR2-dependent activation of PI3K-ERK1/2-NF-κB signaling axis. A, TLR2- or vector-transfected HEK-293 cells were treated with PE_PGRS11. The cells were further stained with antibody specific to PE_PGRS11 followed by Cy5-labeled secondary antibody. The immunofluorescence staining of cells was analyzed by confocal microscopy. B, HEK-293 cells, transiently transfected with vector or TLR2 cDNA construct along with NF-κB reporter construct, were treated with different doses of PE_PGRS11 protein followed by analysis of NF-κB reporter activity (mean ± S.E., n = 3). C, A549 cells were treated with PE_PGRS11 protein for the indicated time points followed by immunoblotting of nuclear and cytosolic fractions of cells for NF-κB. D, activation of p85 (PI3K), ERK1/2, and 4EBP1 by PE_PGRS11. E, transfection of cells with TLR2 dominant-negative (D/N) cDNA construct abrogates PE_PGRS11-induced activation of ERK1/2 and 4EBP1. F, A549 cells were pretreated with LY294002 or AKT inhibitor (Inhi.) followed by analysis of PE_PGRS11-induced activation of ERK1/2. The results are representative of two independent experiments. PCNA, proliferating cell nuclear antigen; p, phospho; Med, medium.