Fig. 1

Induction of IDO1 mRNA and protein levels in lung adenocarcinoma cell lines stimulated with IL-1β. A Heatmap showing the FPKM values of TDO2, IDO1, IDO2, CD274 and RelA mRNA of the cell lines used in this study. B Schematic of experimental set-up illustrating the cell lines tested. C Lung adenocarcinoma cells were treated with 5 ng/mL IL-1β, and cell viability measured 48 h after stimulation (n = 3 biological replicates). D HBEC-3KT and HSAEC-1KT “normal” lung epithelial cells or lung adenocarcinoma cell lines were grown in medium with vehicle control or 5 ng/mL IL-1β for 48 h, and RT-qPCR of IDO1 was performed. Note split scale for IDO1. E RT-qPCR showing induction of CXCL8 mRNA as a measure of activation of IL-1 signaling in HBEC-3KT and HSAEC-1KT cells in response to 5 ng/ml IL-1β treatment for 48 h. F HBEC-3KT and HSAEC-1KT cells show an induction of IDO1 mRNA in response to 5 ng/mL Ifnγ treatment for 48 h. G TDO2 mRNA levels measured by RT-qPCR in normal and lung adenocarcinoma cells in response to IL-1β treatment for 48 h. H IDO1 mRNA levels were measured by RT-qPCR in HCC827 cells treated with 0.1, 1, or 10 ng/mL of IL-1β for 48 h. I RT-qPCR of IDO1 mRNA in H2228 cells treated with 5 ng/mL IL-1β or 100 ng/mL IL-1Ra for 48 h. J Western blot analysis was performed for IDO1 and TDO2 protein levels in H1792 and HCC827 lung adenocarcinoma cells treated with vehicle control or 5 ng/mL IL-1β for 48 h. GAPDH was used as the loading control. K Cell counts were measured for monolayer cultures of H1792 and HCC827 cells stimulated with 5 ng/mL IL-1β ± 20 nM epacadostat (IDOi) for 4 days. Error bars represent ± SD of 3 biological replicates; *P ≤ 0.05, ** ≤ 0.005, *** ≤ 0.0005. mRNA levels were normalized to 18s mRNA and represented as fold change over HBEC-3KT control treatment. GAPDH was used as the loading control