Fig. 3

Effect of GLS-1027 on DC’s differentiation and maturation. (A) Fresh human monocytes were allowed to attach to cell culture plates as described in Materials and Methods and stimulated with GM-CSF/IL-4 for 6 days were allowed to adhere in the absence or presence of GLS-1027 (10µM) for 7 days to develop into dendritic cells. On day 7, the cells were harvested, and were stained with indicated mAbs and analyzed by flow cytometry. Histograms show the staining of specific surface markers, and filled grey histograms represent the isotype-matched control antibody staining, blue line is LPS treatment, and red line is LPS + GLS-1027 treatment. The results are representative of two independent experiments. (B) GLS-1027 inhibits LPS-induced cytokine production of DCs. On day 7 cell culture supernatants from DCs cultured with LPS in the presence of GLS-1027 were measured via ELISA for the concentrations of the cytokines TNF-α, IL-1β, IL-6 and IL-23 in the media. A non-parametric two-tailed t-test (Mann–Whitney) was used for statistical analysis, and the relevant p values are indicated. Error bars in all panels indicate 1 standard deviation among 3 replicates per condition. The experiment was repeated twice with similar results are shown