Soluble TLR4 can block Ni2+/Co2+‐induced IL‐8 production.

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Soluble TLR4 can block Ni2+/Co2+‐induced IL‐8 production. Soluble TLR4 can block Ni2+/Co2+‐induced IL‐8 production. (A) Western blot analysis of the indicated HA‐tagged TLR4 proteins in supernatants and total lysates from transfected HEK293 WT cells. Data demonstrate MD2‐independent release of truncated sTLR4 into the culture supernatant. (B–C) IL‐8 enzyme‐linked immunosorbent assay of HEK293 TLR4/MD2 cells (B) or HUVEC (C) incubated with undiluted supernatants produced by HEK293 WT cells transfected with the indicated constructs. Supernatants were taken 48 h after transfection with the indicated TLR4 constructs and freshly supplemented with diluent (ctrl), Ni2+, Co2+ or LPS before addition to the respective recipient cells and incubated for 8 h. Data represent means±s.d. of three independent experiments. **P<0.01, t‐test. HA, haemagglutinin; HUVEC, human primary umbilical vein endothelial cells; IL, interleukin; LPS, lipopolysaccharide; TLR4, Toll‐like receptor 4; WT, wild‐type. Badrinarayanan Raghavan et al. EMBO Rep. 2012;13:1109-1115 © as stated in the article, figure or figure legend