Figure 5.

The effect of metabolites 1–3 on the expression of pro-inflammatory genes in primary thioglacolate-elicited macrophages post LPS stimulation for 12 h and simultaneous stimulation with the respective disulfide. (A) NO release in cell culture supernatant was measured using Griess reaction, (B) iNOS, (C) IL-6 and (D) IL-12 mRNA levels were quantified using real-time PCR. All treatments have been compared to carbowax 400 0.1% v/v + 0.01% ethanol treated cells in each time point indicated. Disulfide concentration used for 1 and 2 treatments was 15.62 μΜ and for 3 it was 31.25 μΜ. Statistical analysis was carried out using a Mann–Whitney unpaired t-test in Graphpad Prism 7.0 and graphs represent mean ± SEM (* indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001 compared to carbowax 12 h).