Fig. 7.

Determination of GM-CSF/IFN-γ-induced inflammatory tolerance mediated by SOCS3. A, B Western blots showing that treatment with IFN-γ or GM-CSF recombinant protein induces the expression of endogenous SOCS1 and/or SOCS3 in macrophages following treatment at different concentrations for 0.5 h. C, E, G Western blots of pSTAT1/3/5 following different treatments. pi indicates GM-CSF or IFN-γ pretreatment for 0.5 h without a change of culture medium, followed by the addition of the other cytokine for 1.5 h; p indicates pretreatment with GM-CSF or IFN-γ for 0.5 h with a change of medium, followed by the other cytokine for 1.5 h. D, F, H Statistical analysis of data as in C, E, and G, respectively. I–K ELISA assays of TNF-α and IL-1β in the supernatant and lysate of cell cultures after various treatments. Data are expressed as the mean ± SEM; *P < 0.05.