Hsp70 decreases the production of TNF-α and IL-6, but not that of IL-10, in response to TLR agonists. Monocytes were left untreated (NS) or cultured for 48 h in the presence of LPS (100 ng/ml), flagellin (10 μg/ml), peptidoglycan (PG) (10 μg/ml) and porins (3 μg/ml), alone or in the presence of Hsp70 (3 μg/ml). TLR agonists were pre-incubated with Hsp70 in 200 μl of binding buffer for 2 h at 37°C, before adding them to the corresponding wells. The concentrations of TNF-α, IL-6 and IL-10 were measured in the supernatants at the indicated time periods. Two-way ANOVA with Bonferroni post-test: *P < 0.05; **P < 0.01; ***P < 0.001 each TLR agonist vs. each TLR agonist + Hsp70. NS = non-stimulated cells. Data is representative of 3 independent assays.