Figure 2

The effect of histatin 3 on NF-κB-dependent activation by HSC70 through TLR2 and TLR4. (A, B) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR2 and TLR4 in the presence of histatin 3. 293-TLR4/MD2-CD14 (A) and 293-TLR2/CD14 (B) cells were transfected with pIgκB-Luc, and were stimulated with mixtures of the indicated peptides and LPS, HSP70, or HSC70 for 6 h. Cell lysates prepared from the stimulated cells were analyzed by luciferase assays. The values are shown as fold induction of the standardized luciferase activity over the unstimulated control (none). **, P < 0.01; ***, P < 0.001 versus stimulation with HSC70 alone. cont. pep., control peptide. (C, D) Luciferase assays for NF-κB-dependent activation by HSC70 through TLR2 and TLR4 in the presence of histatin family members. The procedures were same as in (A and B), except that stimulations were with mixtures of HSC70 and histatins 3, 4, or 5. Bars represent the means and range of duplicate samples. *, P < 0.05; **, P < 0.01 versus stimulation with HSC70 alone. †, P < 0.05; ††, P < 0.01.