Fig. 4.

Recovered NF-κB signaling in the costimulated cells when SOCS-1 was abolished (A) RAW264.7 cells were transfected with siRNA targeting SOCS-1 mRNA or scrambled siRNA and stimulated with either an agonist for TLR7 or TLR9 or both for indicated time points. The whole cell lysates were extracted, and Western blot analysis showed the cellular levels of IRAK-1, IKK-β, IκBα and SOCS-1. α-tubulin was used as a loading control. (B) RAW264.7 cells were stimulated with each agonist for TLR7 and TLR9 or both for indicated time points, and then nuclear extracts were immunoprecipitated with anti-phospho-NF-κB p65 or anti-SOCS-1, followed by western blot analysis.