Fig. 4

Effects of MSO on protein synthesis and on LPS-triggered production of the chemokine CXCL1. (a, b) mouse peritoneal macrophages were treated as described in Fig. 1 in medium containing 50 μCi of a mixture of S³⁵-labeled methionine and cysteine. At the indicated times after LPS treatment, (a) cells were harvested, TCA-precipitated, filtered and counted, and at the same times (b) supernatant was collected, TCA precipitated, filtered, and counted to determine how overall levels of a, cellular, and b, secreted protein synthesis was affected by MSO treatment shown to reduce production of IL-6 and TNFα. Data shown (s ± SEM) represent 3 macrophage preparations. c, effect of MSO on LPS-triggered production of the cytokine CXCL1 (n = 3). Experiments were carried out as described for Fig. 1 and Fig. 2. ELISA was used to quantitate LPS-triggered production of CXCL1 in the presence or absence of MSO 4 h and 6 h after LPS treatment. CXCL1 was not detectable in the medium of untreated cells, or for cells treated with MSO alone (minus-LPS). The mean CXCL1 protein concentration measured in LPS stimulated controls was 450 pg/mL at 4 h and 750 pg/mL at 6 h. Control values were normalized to 100%, and average values from treated samples were plotted in comparison to those controls