Comprehensive eicosanoid panel.

Eicosanoid profiling was conducted on flash-frozen pancreas tissue from wild type mice (C57B6/J or CD1), KPC mice, or orthotopic tumors (cell lines FC-1199, FC-1242, and FC-1245) grown in C57B6/J mice. In separate experiments, eicosanoid profiling was performed on flash frozen human PDAC or normal pancreas. Age-matched normal and PanIN-bearing pancreata were included from a previous study⁴. Tissues were homogenized in 1 ml of PBS containing 10% ethanol and 300 μl were extracted using strata-x polymeric reverse phase columns (88-S100-UBJ Phenomenex). Samples were taken up in 50 μl of 63% H₂0, 37% ACN, 0.02% Acetic Acid, and 10 μl was injected into UPLC (ACQUITY UPLC System, Waters) and analyzed on a Sciex 6500 Qtrap mass spectrometer at the UCSD Lipidomics Core as previously described³⁷. Tissue eicosanoid concentrations were quantified using deuterated internal standards in conjunction with standard curves obtained in parallel using identical conditions as previously described³⁸, and were normalized to total protein mass of the sample.