FIG 1.
Triggering of endosomal TLR7 and TLR9 but not TLR3 activates NF-κB in S11 cells. (A) Expression of TLR3, TLR7, and TLR9 mRNA was detected in S11 cells by reverse transcription-PCR. (B) Triggering of TLR7 and TLR9 but not TLR3 activates NF-κB in S11 cells. NF-κB subunit p65 was detected in nuclear extracts of S11 cells at different time points after mock treatment or treatment with 25 μg/ml poly(I·C), 3 μM R848, 0.5 μM CpG ODN 1826, or 1 μM NF-κB inhibitor Bay 11-7082 by Western blot analysis. PCNA was used as a loading control. (C) Triggering of TLR3 induces a pattern of cytokine expression different from that seen with TLR7 or TLR9. S11 cells were stimulated with TLR ligands as described for panel B and harvested at different time points. Expression of IFN-β, IL-6, and IL-10 was analyzed by qPCR, normalized to GAPDH, and plotted relative to mock-treated control results. Data shown in panel C are means ± standard deviations (SD) of the results of three independent experiments. Some of the error bars are covered by the symbols.