Figure 6

The role of TLR2/TLR1 during Nm -mediated gene activation. (A) Semi-quantitative RT-PCR was performed to measure activation of nfkbiz, il6, and il8 after 4 h treatment with either α14, MC58, MC58porB^-, or MC58siaD^- Nm strains at a multiplicity of infection (MOI) of 10, respectively. Control experiments were performed in absence of a stimulus. PCR reactions were analyzed after the cycle numbers indicated at the top of the panels. (B) Semi-quantitative RT-PCR was performed to measure activation of nfkbiz, il6, and il8 after 4 h treatment with either Fsl-1, Nm MC58siaD^-, or PAM3CSK4 at the indicated MOI or concentrations, respectively. Control experiments were performed in absence of a stimulus. PCR reactions were analyzed after the cycle numbers indicated at the top of the panels. (C) HIBCPP cells were challenged with 10 ng mL^-1 PAM3CSK4 or Fsl-1 for the indicated time. Expression of IκBζ protein was analyzed by immunoblot analysis. Detection of β-actin protein levels served as control. (D) Semi-quantitative RT-PCR was performed to determine activation of nfkbiz, il6, or il8 in primary human monocytes after treatment with the indicated amounts of PAM3CSK4 or “ultrapure” LPS. Control experiments were performed in absence of a stimulus. PCR reactions were analyzed after the cycle numbers indicated at the top of the panels.