Additional file 2: of S100A8 and S100A9 proteins form part of a paracrine feedback loop between pancreatic cancer cells and monocytes

Figure S1. Specificity of signalling downstream of S100A8 and S100A9. Panc-1 cells were left untreated or incubated in the presence of polymyxin-b (10 μg/mL). The cells were transfected with NF-κB and control reporters [2 μg of DNA] and stimulated with recombinant A8-GST, A9-GST or GST [2 μg/mL] for 24 h. Cells treated with TNF-α [10 ng/mL] served as a positive control for NF-κB induction. For blocking conditions, cells were treated with anti-RAGE blocking antibody for 1 h prior to stimulation. S100A8-GST and S100A9-GST were incubated with anti-S100A8 and anti-S100A9 neutralising antibodies for 1 h before addition to cancer cell cultures. Luciferase activity was plotted as the mean ± SEM of three independent experiments performed in triplicate. Error bars represent standard error (* P