Figure 1. IDO mRNA was quantified by real-time PCR. A: Products show the results from RNA extracted from four pooled corneal endothelial cell cultures. IDO mRNA levels are normalized to GAPDH. Higher levels of IDO mRNA expression are seen in pools of HCECs that have been stimulated with 500 ng/ml IFN-γ and 500 ng/ml IFN-γ+100 U/ml TNF-α for three days. The y-axis indicates IDO mRNA fold change. The x-axis presents the difference in four pool treatments. Lane 1, unstimulated; lane 2, 500 ng/ml IFN-γ; lane 3, 100 U/ml TNF-α; lane 4, 500 ng/ml IFN-γ + 100 U/ml TNF-α. B: The bottom panels show examples of the determination of RNA using the Bioanalyzer: unstimulated HCECs (top), IFN-γ treated HCECs (middle), and IFN-γ/TNF-α treated HCECs (bottom).