Lipidomic Analysis of Dynamic Eicosanoid Responses During the Induction and Resolution of Lyme Arthritis
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1 Lipidomic Analysis of Dynamic Eicosanoid Responses During the Induction and Resolution of Lyme Arthritis Victoria A. Blaho 1, Matthew W. Buczynski 2, Charles R. Brown 1, and Edward A. Dennis 2 1 Department of Veterinary Pathobiology, University of Missouri, 315 Connaway Hall, Columbia, MO Department of Pharmacology and Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive, La Jolla, CA These authors contributed equally to this manuscript. Running title: Eicosanoid lipidomics of Lyme arthritis Address correspondence to: Charles R. Brown, University of Missouri, 315 Connaway Hall, Columbia, MO 65211, BrownChar@missouri.edu or Edward A. Dennis, University of California San Diego, University of California, San Diego, 9500 Gilman Dr., La Jolla, CA , edennis@ucsd.edu. Supplemental Figures S1-S3 Supplemental Tables 1-5 1
2 Day Sham C3H (Susceptible) PG Synthase AA DGLA EPA AdA 6k PGF1α TxB2 PGF2α PGE2 PGD2 11β PGF2α TXB1 PGF1α PGE1 PGD1 17 6k PGF1α TXB3 PGF3α PGE3 PGD3 dihomo PGF2α dihomo PGE2 dihomo PGD2 dihomo PGJ2 dihomo 15d PGD2 COX 5-LOX CYP PGDH CYP β-ox non enzymatic LTAH LTCS 15-LOX 12-LOX HEDH seh 6k PGE1 6,15 dk-,dh- PGF1α 15k PGF1α 15k PGF2α 15k PGE2 dh PGF2α dhk PGF2a dhk PGE2 dhk PGD2 bicyclo PGE2 11β dhk PGF2α 19oh PGF2α 20oh PGF2α 19oh PGE2 20oh PGE2 2,3 dinor 11β PGF2α PGFM PGEM 11β PGE2 PGK2 12-HHT 11-HETE PGA2 PGB2 15d PGA2 PGJ2 15d PGD2 15d PGJ2 5-iso PGF2a VI 8-iso PGF2a III 9-HETE LTB4 20oh LTB4 20cooh LTB4 5,6-diHETE Δ6t LTB4 12epi LTB4 Δ6t,12epi LTB4 LTC4 LTD4 LTE4 Δ11t LTC4 Δ11t LTD4 Δ11t LTE4 5-HETE 5,15-diHETE 6R-LXA4 LXA4 LXB4 RvE1 RvD1 PD1 Δ15t-PD1 10S,17S-DiHDoHE 8,15-diHETE 15-HETE 17-HDoHE 13-HODE 8-HETE EXC4 EXD4 EXE4 12-HETE 9-HODE HXA3 HXB3 5-oxoETE 12-oxoETE 15-oxoETE 9-oxoODE 13-oxoODE 20-HETE 5,6-EET 8,9-EET 11,12-EET 14,15-EET 9,10 EpOME 12,13 EpOME 5,6-DHET 8,9-DHET 11,12-DHET 14,15-DHET 9,10 dihome 12,13 dihome Decrease Increase ND
3 A EET/ DHET ratio ,6 8,9 11,12 14,15 B Day post-infection Day post-infection 3
4 A pg/mg tissue Δ15t-PD 1 #, #, C3H DBA B # 10S,17S-DiHDoHE #, C3H DBA Day post-infection Day post-infection 4
5 Supplemental Tables 1-5 Supplemental Table 1. Liquid Chromatography and Mass Spectrometry Parameters. Period Eicosanoid LIPID MAPS ID 1 2 Precursor Ion Product Ion Retention Time Limit of Detection (pg) Internal Standard PGFM LMFA (d 4 ) PGF 2α PGEM LMFA (d 4 ) PGE 2 20oh PGF 2α LMFA (d 4 ) PGF 2α 19oh PGF 2α LMFA (d 4 ) PGF 2α 20oh PGE 2 LMFA (d 4 ) PGE 2 19oh PGE 2 LMFA (d 4 ) PGE k PGF 1α LMFA (d 4 ) 6k PGF 1α 20cooh LTB 4 LMFA (d 4 ) LTB 4 6k PGF 1α LMFA (d 4 ) 6k PGF 1α (d 4 ) 6k PGF 1α LMFA RvE 1 LMFA (d 4 ) LTB 4 2,3 dinor 11β PGF 2α LMFA (d 4 ) PGF 2α 20oh LTB 4 LMFA (d 4 ) LTB 4 6k PGE 1 LMFA (d 4 ) 6k PGF 1α TXB 3 LMFA (d 4 ) TXB 2 TXB 1 LMFA (d 4 ) TXB 2 PGF 3α LMFA (d 4 ) PGF 2α (d 4 ) TXB 2 LMFA TXB 2 LMFA (d 4 ) TXB 2 6,15 dk-,dh- PGF 1α LMFA (d 4 ) PGF 2α PGE 3 LMFA (d 4 ) PGE 2 8-iso PGF 2α III LMFA (d 11 ) 5-iso PGF 2α VI 11β PGF 2α LMFA (d 4 ) PGF 2α PGD 3 LMFA (d 4 ) PGD 2 (d 11 ) 5-iso PGF 2α VI LMFA iso PGF 2α VI LMFA (d 11 ) 5-iso PGF 2α VI (d 4 ) PGF 2α LMFA PGF 2α LMFA (d 4 ) PGF 2α PGF 1α LMFA (d 4 ) PGF 2α (d 4 ) PGE 2 LMFA PGE 2 LMFA (d 4 ) PGE 2 PGK 2 LMFA (d 4 ) PGE 2 11β PGE 2 LMFA (d 4 ) PGE 2 LXB 4 LMFA (d 4 ) LTB 4 PGE 1 LMFA (d 4 ) PGE 2 EXC 4 LMFA (d 4 ) LTB 4 15k PGF 2α LMFA (d 4 ) PGF 2α (d 4 ) PGD 2 LMFA PGD 2 LMFA (d 4 ) PGD 2 PGD 1 LMFA (d 4 ) PGD 2 15k PGE 2 LMFA (d 4 ) PGE 2 15k PGF 1α LMFA (d 4 ) PGF 2α dh PGF 2α LMFA (d 4 ) PGF 2α EXD 4 LMFA (d 4 ) LTB 4 11β dhk PGF 2α LMFA (d 4 ) PGF 2α RvD 1 LMFA (d 4 ) LTB 4 6R-LXA 4 LMFA (d 4 ) LTB 4 dhk PGE 2 LMFA (d 4 ) dhk PGD 2 (d 4 ) dhk PGF 2α LMFA dhk PGF 2α LMFA (d 4 ) dhk PGF 2α LXA 4 LMFA (d 4 ) LTB 4 5
6 Supplemental Table 1. Continued. Period Eicosanoid LIPID MAPS ID 3 4 Precursor Ion Product Ion Retention Time Limit of Detection (pg) Internal Standard EXE 4 LMFA (d 4 ) LTB 4 dihomo PGF 2α LMFA (d 4 ) PGF 2α (d 4 ) dhk PGD 2 LMFA dhk PGD 2 LMFA (d 4 ) dhk PGD 2 dihomo PGE 2 LMFA (d 4 ) PGE 2 LTC 4 LMFA (d 4 ) LTB 4 dihomo PGD 2 LMFA (d 4 ) PGD 2 PGA 2 LMFA (d 4 ) 15d PGJ 2 LTD 4 LMFA (d 4 ) LTB 4 PGJ 2 LMFA (d 4 ) 15d PGJ 2 PGB 2 LMFA (d 4 ) 15d PGJ 2 11 LTC 4 LMFA (d 4 ) LTB 4 LTE 4 LMFA (d 4 ) LTB 4 11 LTD 4 LMFA (d 4 ) LTB 4 PD 1 LMFA (d 4 ) LTB 4 bicyclo PGE 2 LMFA (d 4 ) PGE 2 15t-PD 1 LMFA (d 4 ) LTB 4 11 LTE 4 LMFA (d 4 ) LTB 4 8,15-diHETE LMFA (d 4 ) LTB 4 10S,17S-DiHDoHE LMFA (d 4 ) LTB 4 5,15-diHETE LMFA (d 4 ) LTB 4 6t LTB 4 LMFA (d 4 ) LTB 4 12epi LTB 4 LMFA (d 4 ) LTB 4 6t,12epi LTB 4 LMFA (d 4 ) LTB 4 (d 4 ) 12,13 dihome LMFA ,13 dihome LMFA (d 4 ) 12,13-diHOME (d 4 ) LTB 4 LMFA d PGD 2 LMFA (d 4 ) 15d PGJ 2 LTB 4 LMFA (d 4 ) LTB 4 dihomo PGJ 2 LMFA (d 4 ) 15d PGJ 2 (d 4 ) 9,10 dihome LMFA ,10 dihome LMFA (d 4 ) 9,10-diHOME 14,15-DHET LMFA (d 8 ) 5-HETE 12-HHT LMFA (d 8 ) 5-HETE dihomo 15d PGD 2 LMFA (d 4 ) 15d PGJ 2 11,12-DHET LMFA (d 8 ) 5-HETE HXA 3 LMFA (d 8 ) 11,12-EET HXB 3 LMFA (d 8 ) 11,12-EET (d 4 ) 15d PGJ 2 LMFA d PGJ 2 LMFA (d 4 ) 15d PGJ 2 8,9-DHET LMFA (d 8 ) 5-HETE (d 6 ) 20-HETE LMFA HETE LMFA (d 6 ) 20-HETE 15d PGA 2 LMFA (d 4 ) 15d PGJ 2 5,6-diHETE LMFA (d 4 ) LTB 4 5,6-DHET LMFA (d 8 ) 5-HETE 6
7 Supplemental Table 1. Continued. Period Eicosanoid LIPID MAPS ID Precursor Product Retention Limit of Detection Ion Ion Time (pg) Internal Standard (d 4 ) 13-HODE LMFA HODE LMFA (d 4 ) 13-HODE (d 4 ) 9-HODE LMFA (d 8 ) 15-HETE LMFA HDoHE LMFA (d 8 ) 15-HETE 9-HODE LMFA (d 4 ) 9-HODE 15-HETE LMFA (d 8 ) 15-HETE 5 13-oxoODE LMFA (d 7 ) 5-oxoETE 15-oxoETE LMFA (d 7 ) 5-oxoETE 11-HETE LMFA (d 8 ) 5-HETE 9-oxoODE LMFA (d 7 ) 5-oxoETE 12-HETE LMFA (d 8 ) 15-HETE 8-HETE LMFA (d 8 ) 5-HETE 12-oxoETE LMFA (d 7 ) 5-oxoETE 9-HETE LMFA (d 8 ) 5-HETE (d 8 ) 5-HETE LMFA HETE LMFA (d 8 ) 5-HETE (d 8 ) 14,15-EET LMFA ,15-EET LMFA (d 8 ) 14,15-EET 12,13-EpOME LMFA (d 8 ) 11,12-EET (d 7 ) 5-oxoETE LMFA oxoETE LMFA (d 7 ) 5-oxoETE 9,10-EpOME LMFA (d 8 ) 11,12-EET 11,12-EET LMFA (d 8 ) 11,12-EET (d 8 ) 8,9-EET LMFA (d 8 ) 11,12-EET LMFA ,9-EET LMFA (d 8 ) 8,9-EET 5,6-EET LMFA (d 8 ) 11,12-EET 7
8 Supplemental Table 2. Eicosanoid levels (pg per mg tissue) in B. burgdorferi-infected C3H mice. Eicosanoid A Day 0 Day 3 Day 7 Day 10 Day 14 Day 17 Day 21 Day 24 Day 28 Day 35 6k PGF 1α 0.8 ± ± ± ± ± ± ± ± ± ± 0.4 TXB ± ± ± ± ± ± ± ± ± ± 0.1 PGE ± ± ± ± ± ± ± ± ± ± 0.1 PGD ± ± ± ± ± ± ± ± ± ± k PGE ± ± ± ± ± ± ± ± ± ± 0.0 dhk PGE ± ± ± ± ± ± ± ± ± ± 0.0 PGJ ± ± ± ± ± ± ± ± ± ± d PGD ± ± ± ± ± ± ± ± ± ± d PGJ ± ± ± ± ± ± ± ± ± ± β PGE ± ± ± ± ± ± ± ± ± ± HHT 0.2 ± ± ± ± ± ± ± ± ± ± HETE 1.4 ± ± ± ± ± ± ± ± ± ± iso PGF 2α VI 0.0 ± ± ± ± ± ± ± ± ± ± iso PGF 2α III 0.0 ± ± ± ± ± ± ± ± ± ± HETE 1.9 ± ± ± ± ± ± ± ± ± ± 0.4 5,6-diHETE 0.0 ± ± ± ± ± ± ± ± ± ± 0.0 LTC ± ± ± ± ± ± ± ± ± ± 0.1 LTE ± ± ± ± ± ± ± ± ± ± HETE 4.6 ± ± ± ± ± ± ± ± ± ± 0.9 RvD ± ± ± ± ± ± ± ± ± ± 0.0 PD ± ± ± ± ± ± ± ± ± ± t-PD ± ± ± ± ± ± ± ± ± ± S,17S-DiHDoHE 0.2 ± ± ± ± ± ± ± ± ± ± HDoHE 5.1 ± ± ± ± ± ± ± ± ± ± HETE 3.6 ± ± ± ± ± ± ± ± ± ± HODE 24.1 ± ± ± ± ± ± ± ± ± ± HETE 1.7 ± ± ± ± ± ± ± ± ± ± 0.9 8,15-diHETE 0.0 ± ± ± ± ± ± ± ± ± ± HODE 26.2 ± ± ± ± ± ± ± ± ± ± HETE 3.0 ± ± ± ± ± ± ± ± ± ± 0.6 HXA ± ± ± ± ± ± ± ± ± ± 0.3 HXB ± ± ± ± ± ± ± ± ± ± oxoETE 1.6 ± ± ± ± ± ± ± ± ± ± oxoETE 0.2 ± ± ± ± ± ± ± ± ± ± oxoETE 0.8 ± ± ± ± ± ± ± ± ± ± oxoODE 3.6 ± ± ± ± ± ± ± ± ± ± oxoODE 1.5 ± ± ± ± ± ± ± ± ± ± 0.5 5,6-EET 0.8 ± ± ± ± ± ± ± ± ± ± 0.2 8,9-EET 0.5 ± ± ± ± ± ± ± ± ± ± ,12-EET 0.3 ± ± ± ± ± ± ± ± ± ± ,15-EET 0.6 ± ± ± ± ± ± ± ± ± ± 0.1 9,10-EpOME 2.4 ± ± ± ± ± ± ± ± ± ± ,13-EpOME 4.2 ± ± ± ± ± ± ± ± ± ± 0.7 5,6-DHET 0.1 ± ± ± ± ± ± ± ± ± ± 0.0 8,9-DHET 0.1 ± ± ± ± ± ± ± ± ± ± ,12-DHET 1.9 ± ± ± ± ± ± ± ± ± ± ,15-DHET 0.2 ± ± ± ± ± ± ± ± ± ± 0.0 9,10-diHOME 1.8 ± ± ± ± ± ± ± ± ± ± ,13-diHOME 3.3 ± ± ± ± ± ± ± ± ± ± 0.5 Metabolites are listed in the same order as they are displayed on the heat map in Figure 2 of the text. Values are the mean ± s.e.m. of 8-10 mice at each time point. A Eicosanoids listed were detected in at least one sample at one time point. Values were rounded to one decimal point; thus some metabolites are shown with values of 0.0±0.0 at every time point. All eicosanoids from Supplemental Table 5 that are unlisted in the current table were measured but not detected. 8
9 Supplemental Table 3. Eicosanoid levels (pg per mg tissue) in B. burgdorferi-infected DBA mice. Eicosanoid A Day 0 Day 3 Day 7 Day 10 Day 14 Day 17 Day 21 Day 24 Day 28 B Day 35 6k PGF 1α 1.2 ± ± ± ± ± ± ± ± ± ± 0.2 TXB ± ± ± ± ± ± ± ± ± ± 0.0 PGF 2α 0.0 ± ± ± ± ± ± ± ± ± ± 0.1 PGE ± ± ± ± ± ± ± ± ± ± 0.1 PGD ± ± ± ± ± ± ± ± ± ± k PGE ± ± ± ± ± ± ± ± ± ± 0.0 dhk PGD ± ± ± ± ± ± ± ± ± ± d PGD ± ± ± ± ± ± ± ± ± ± d PGJ ± ± ± ± ± ± ± ± ± ± β PGE ± ± ± ± ± ± ± ± ± ± HHT 1.4 ± ± ± ± ± ± ± ± ± ± HETE 1.5 ± ± ± ± ± ± ± ± ± ± iso PGF 2α VI 0.2 ± ± ± ± ± ± ± ± ± ± HETE 1.2 ± ± ± ± ± ± ± ± ± ± 0.1 LTC ± ± ± ± ± ± ± ± ± ± 0.0 LTE ± ± ± ± ± ± ± ± ± ± HETE 5.3 ± ± ± ± ± ± ± ± ± ± 0.7 6S-LXA ± ± ± ± ± ± ± ± ± ± 0.0 RvD ± ± ± ± ± ± ± ± ± ± 0.1 PD ± ± ± ± ± ± ± ± ± ± t-PD ± ± ± ± ± ± ± ± ± ± S,17S-DiHDoHE 0.4 ± ± ± ± ± ± ± ± ± ± HDoHE 7.6 ± ± ± ± ± ± ± ± ± ± HETE 5.7 ± ± ± ± ± ± ± ± ± ± HODE 58.5 ± ± ± ± ± ± ± ± ± ± HETE 2.3 ± ± ± ± ± ± ± ± ± ± HODE 49.4 ± ± ± ± ± ± ± ± ± ± HETE 6.6 ± ± ± ± ± ± ± ± ± ± 0.7 HXA ± ± ± ± ± ± ± ± ± ± 0.4 HXB ± ± ± ± ± ± ± ± ± ± oxoETE 2.2 ± ± ± ± ± ± ± ± ± ± oxoETE 8.8 ± ± ± ± ± ± ± ± ± ± oxoETE 1.6 ± ± ± ± ± ± ± ± ± ± oxoODE 11.4 ± ± ± ± ± ± ± ± ± ± oxoODE 9.4 ± ± ± ± ± ± ± ± ± ± 0.8 5,6-EET 4.0 ± ± ± ± ± ± ± ± ± ± 0.8 8,9-EET 1.9 ± ± ± ± ± ± ± ± ± ± ,12-EET 1.1 ± ± ± ± ± ± ± ± ± ± ,15-EET 1.3 ± ± ± ± ± ± ± ± ± ± 0.1 9,10-EpOME 8.6 ± ± ± ± ± ± ± ± ± ± ,13-EpOME 8.7 ± ± ± ± ± ± ± ± ± ± 0.5 5,6-DHET 0.2 ± ± ± ± ± ± ± ± ± ± 0.0 8,9-DHET 0.1 ± ± ± ± ± ± ± ± ± ± ,12-DHET 1.9 ± ± ± ± ± ± ± ± ± ± ,15-DHET 0.2 ± ± ± ± ± ± ± ± ± ± 0.0 9,10-diHOME 2.5 ± ± ± ± ± ± ± ± ± ± ,13-diHOME 4.1 ± ± ± ± ± ± ± ± ± ± 0.5 Metabolites are listed in the same order as they are displayed on the heat map in Figure 2 of the text. Values are the mean ± s.e.m. of 8-10 mice at each time point. A Eicosanoids listed were detected in at least one sample at one time point. Values were rounded to one decimal point; thus some metabolites are shown with values of 0.0±0.0 at every time point. All eicosanoids from Supplemental Table 5 that are unlisted in the current table were measured but not detected. B n = 5 mice. 9
10 Supplemental Table 4. Eicosanoid levels (pg per mg tissue) in C3H sham-infected mice. Eicosanoid A Day 0 Day 3 Day 7 Day 10 Day 14 Day 17 Day 21 Day 24 Day 28 Day 35 6k PGF 1α 0.2 ± ± ± ± ± ± ± ± ± ± 0.0 TXB ± ± ± ± ± ± ± ± ± ± 0.1 PGF 2α 0.1 ± ± ± ± ± ± ± ± ± ± 0.0 PGE ± ± ± ± ± ± ± ± ± ± 0.0 PGD ± ± ± ± ± ± ± ± ± ± 0.0 dhk PGD ± ± ± ± ± ± ± ± ± ± 0.0 PGJ ± ± ± ± ± ± ± ± ± ± d PGD ± ± ± ± ± ± ± ± ± ± d PGJ ± ± ± ± ± ± ± ± ± ± β PGE ± ± ± ± ± ± ± ± ± ± HHT 0.0 ± ± ± ± ± ± ± ± ± ± HETE 1.2 ± ± ± ± ± ± ± ± ± ± HETE 0.8 ± ± ± ± ± ± ± ± ± ± 0.1 LTE ± ± ± ± ± ± ± ± ± ± HETE 2.2 ± ± ± ± ± ± ± ± ± ± 0.6 RvD ± ± ± ± ± ± ± ± ± ± 0.0 PD ± ± ± ± ± ± ± ± ± ± S,17S-DiHDoHE 0.0 ± ± ± ± ± ± ± ± ± ± HDoHE 1.4 ± ± ± ± ± ± ± ± ± ± HETE 1.8 ± ± ± ± ± ± ± ± ± ± HODE 10.2 ± ± ± ± ± ± ± ± ± ± HETE 0.9 ± ± ± ± ± ± ± ± ± ± HODE 10.2 ± ± ± ± ± ± ± ± ± ± HETE 1.6 ± ± ± ± ± ± ± ± ± ± 0.1 HXA ± ± ± ± ± ± ± ± ± ± 0.2 HXB ± ± ± ± ± ± ± ± ± ± oxoETE 0.8 ± ± ± ± ± ± ± ± ± ± oxoETE 21.5 ± ± ± ± ± ± ± ± ± ± oxoETE 0.6 ± ± ± ± ± ± ± ± ± ± oxoODE 2.3 ± ± ± ± ± ± ± ± ± ± oxoODE 1.4 ± ± ± ± ± ± ± ± ± ± HETE 0.1 ± ± ± ± ± ± ± ± ± ± 0.0 5,6-EET 1.8 ± ± ± ± ± ± ± ± ± ± 0.1 8,9-EET 0.3 ± ± ± ± ± ± ± ± ± ± ,12-EET 0.2 ± ± ± ± ± ± ± ± ± ± ,15-EET 0.5 ± ± ± ± ± ± ± ± ± ± 0.0 9,10-EpOME 1.2 ± ± ± ± ± ± ± ± ± ± ,13-EpOME 1.9 ± ± ± ± ± ± ± ± ± ± 0.3 5,6-DHET 0.1 ± ± ± ± ± ± ± ± ± ± 0.0 8,9-DHET 0.2 ± ± ± ± ± ± ± ± ± ± ,12-DHET 2.2 ± ± ± ± ± ± ± ± ± ± ,15-DHET 0.2 ± ± ± ± ± ± ± ± ± ± 0.0 9,10-diHOME 1.4 ± ± ± ± ± ± ± ± ± ± ,13-diHOME 3.7 ± ± ± ± ± ± ± ± ± ± 0.3 Metabolites are listed in the same order as they are displayed on the heat map in Supplemental Figure 1 of the text. Values are the mean ± s.e.m. of 6 mice at each time point. A Eicosanoids listed were detected in at least one sample at one time point. Values were rounded to one decimal point; thus some metabolites are shown with values of 0.0±0.0 at every time point. All eicosanoids from Supplemental Table 5 that are unlisted in the current table were measured but not detected. 10
11 Supplemental Table 5. Day 10 eicosanoid levels (pg per mg tissue) in joints of B. burgdorferi-infected C3H COX-2 knockout mice and wild-type controls. Eicosanoid A wild-type COX-2 KO 6k PGF 1α 18.5 ± ± 1.1 TXB ± ± 0.6 PGF 2α 2.3 ± ± 0.2 PGE ± ± 0.4 PGD ± ± k PGE ± ± 0.0 dhk PGD ± ± 0.0 PGJ ± ± d PGD ± ± d PGJ ± ± HHT 4.2 ± ± HETE 5.9 ± ± iso PGF 2α VI 0.1 ± ± iso PGF 2α III 0.1 ± ± HETE 2.2 ± ± 0.3 LTB ± ± 0.0 5,6-diHETE 4.7 ± ± 0.0 LTC ± ± 0.1 LTE ± ± HETE 11.3 ± ± 1.1 PD ± ± t-PD ± ± S,17S-DiHDoHE 0.2 ± ± HDoHE 4.2 ± ± HETE 12.0 ± ± HODE 25.6 ± ± HETE 2.3 ± ± 0.3 8,15-diHETE 0.0 ± ± HODE 0.4 ± ± HETE 9.7 ± ± 2.1 HXA ± ± 2.2 HXB ± ± oxoETE 4.8 ± ± oxoETE 3.2 ± ± oxoETE 4.2 ± ± oxoODE 7.0 ± ± oxoODE 4.8 ± ± HETE 0.4 ± ± 0.0 8,9-EET 1.5 ± ± ,12-EET 0.8 ± ± ,15-EET 1.4 ± ± 0.1 9,10-EpOME 3.5 ± ± ,13-EpOME 5.8 ± ± 0.9 5,6-DHET 0.2 ± ± 0.0 8,9-DHET 0.1 ± ± ,12-DHET 2.1 ± ± ,15-DHET 0.3 ± ± 0.0 9,10-diHOME 2.5 ± ± ,13-diHOME 6.1 ± ± 0.9 Metabolites are listed in the same order as they are displayed on the heat map in Figure 6 of the text. Values are the mean ± s.e.m. of 4 mice at each time point. A Eicosanoids listed were detected in at least one sample at one time point. Values were rounded to one decimal point; thus some metabolites are shown with values of 0.0±0.0 at both time points. All eicosanoids from Supplemental Table 5 that are unlisted in the current table were measured but not detected. 11
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