Figure 4

TLR2/6 mediated signalling using HEK293T cells. HEK293T cells were transfected with pDUO-hTLR6/TLR2 (Invivogen) alone (TLR2/6 (−)) or together with the reporter plasmid pNiFty2-SEAP (Invivogen). Cells were transiently transfected with the respective plasmids as described in Methods. 24h after transfection, cells were incubated with Pam2CSK4 (50 ng/ml), LTA at 0.1, 1 and 10 μg/ml, mLTA1 (10 μg/ml), mLTA2 (10 μg/ml), LGG wild-type or dltD mutant cells (10⁸ CFU/ml) for 24 hours. After incubation, NF-κB-induced SEAP activity was assessed using pNPP (p-nitrophynyl phosphate) and measuring the OD at 405nm. Data represent mean values ± SD. ^*p-value < 0.05 compared to the negative control (TLR2/6(−)).