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FIGURE LEGENDS 715
Fig.1 Main changes produced by lipoic acid supplementation in esterification of [1-14C] 18:3n-716
3 and its products into phospholipids (A) and neutral lipids (B). Data are shown as mean ± 717
SEM (n=3).
718
Fig. 2 Main changes produced by increasing dietary levels of EPA and/or DHA in 719
esterification of [1-14C] 18:3n-3 and its products into phospholipids (A) and neutral lipids (B).
720
Data are shown as mean ± SEM (n=6).
721
Fig. 3 Total radiolabeled docosahexaenoic acid (DHA; 22:6n-3) esterified into cellular lipids 722
(PL+NL) in hepatocytes incubated with [1-14C] 18:3n-3 in the presence or absence of lipoic 723
acid. Cells were isolated from fish fed diets containing different levels of EPA and/or DHA for 724
26 weeks before the experiment. Data are shown as mean ± SEM (n=3).
725
Fig. 4 Relative changes in mRNA transcript abundance of genes involved in the n-3 fatty 726
acid biosynthetic pathway. Atlantic salmon hepatocytes cultivated in the presence or 727
absence of lipoic acid were isolated from fish fed 10 experimental diets containing different 728
levels of EPA and/or DHA for 26 weeks. Samples (n=3) were analyzed using real-time qPCR 729
and data are presented as -∆∆Ct ± SEM. Cells isolated from fish fed the 0 % diet were used 730
as control and values were set to zero. Results are compared by two-way analysis of 731
variance (diet and lipoic acid as factors; P<0.05).
732
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23 TABLES
733
Table 1. Fatty acid composition (mol%) in the experimental diets.
734
0% 0.5% EPA 1.0% EPA 2.0% EPA 0.5% DHA 1.0% DHA 2.0% DHA 0.5% EPA+DHA 1.0% EPA+DHA 2.0% EPA+DHA
16:0 17.7 17.2 16.8 15.9 17.3 16.9 16.0 17.2 16.8 16.0
18:0 4.2 4.1 4.0 3.9 4.2 4.2 4.2 4.2 4.1 4.1
SFA1 23.0 22.4 21.9 20.7 22.3 22.3 21.3 22.5 22.1 21.1
18:1n-9 44.5 43.4 41.7 38.3 43.5 42.0 39.4 43.1 41.8 39.0
MUFA2 47.3 45.8 44.2 41.0 46.3 44.9 42.4 45.8 44.7 41.6
18:2n-6 23.7 23.0 22.5 21.3 23.2 22.8 21.5 23.1 22.5 21.5
18:3n-3 5.3 5.1 4.9 4.9 5.1 4.9 4.6 5.1 5.0 4.6
C18 PUFA3 29.0 28.3 27.7 26.5 28.5 27.8 26.3 28.4 27.6 26.3
20:5n-3 0.0 2.2 4.3 8.4 0.4 0.7 1.4 1.4 2.5 5.1
22:6n-3 0.1 0.6 1.1 1.9 1.8 3.6 7.4 1.2 2.3 4.7
LC-PUFA4 0.7 3.5 6.3 11.8 2.9 5.0 10.1 3.4 5.6 11.0
1Includes 14:0, 20:0, 22:0, 24:0.
735 2
Includes 20:1n-9, 20:1n-11, 22:1n-11.
736
3Includes 18:3n-6.
737
4Includes 20:2n-6, 20:3n-6, 20:4n-6, 22:5n-3.
738 739
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24 740
Table 12. Atlantic salmon primer sequences used for real-time PCR.
741
Gene Accession no. Direction Primer sequence 5´→→→→ 3´
ef1α AF321836 Forward CACCACCGGCCATCTGATCTACAA Reverse TCAGCAGCCTCCTTCTGAACTTC etif3 DW542195 Forward CAGGATGTTGTTGCTGGATGGG
Reverse ACCCAACTGGGCAGGTCAAGA rpol2 CA049789 Forward TAACGCCTGCCTCTTCACGTTGA
Reverse ATGAGGGACCTTGTAGCCAGCAA aco DQ364432 Forward CCTTCATTGTACCTCTCCGCA
Reverse CATTTCAACCTCATCAAAGCCAA
∆5fad AF478472 Forward GCTTGAGCCCGATGGAGG Reverse CAAGATGGAATGCGGAAAATG
∆6fad_a AY458652 Forward TCCCCAGACGTTTGTGTCAGATGC Reverse GCTTTGGATCCCCCATTAGTTCCTG elovl2 TC91192 Forward CGGGTACAAAATGTGCTGGT
Reverse TCTGTTTGCCGATAGCCATT elovl5b NM_001136552 Forward GCAACCTTGACCCAAACAGG
Reverse CCTTGTCTCTACGCAAGGGA srebp1 NM_001195818 Forward AGCTGCACGGCTTCCAGCAG
Reverse TCCTCCGTCTTGGCTCCGGG
Elongation factor 1 alpha (ef1α) eukaryotic translation initiation factor 3 (etif3), RNA polymerase II polypeptide (rpol2), acyl-CoA oxidase (aco), ∆5 desaturase (∆5fad), ∆6
742
desaturase isoform a (∆6fad_a), elongase 2 (elovl2), elongase 5b (elovl5b), sterol regulatory element binding protein 1 (srebp1).
743
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25
Table 23. Fatty acid composition (mol%) in the PL and NL fraction of hepatocytes from Atlantic salmon fed the experimental diets for 26 weeks 744
For Peer Review
26
PUFA3 21.9 ± 2.03b 35.7 ± 5.59ab 36.4 ± 3.34ab 42.0 ± 7.06ab 36.9 ± 1.37ab 38.5 ± 3.17ab 43.7 ± 1.20a 32.8 ± 6.50ab 41.7 ± 4.26ab 49.2 ± 1.95a 0.017 n-6 PUFA 14.4 ± 1.41 18.3 ± 1.28 17.6 ± 1.40 17.2 ± 1.32 20.1 ± 0.29 20.4 ± 0.54 19.3 ± 0.39 18.4 ± 2.67 19.1 ± 0.28 17.9 ± 0.54 0.12 n-6/n-3 2.9 ± 0.82a 1.4 ± 0.24ab 1.1 ± 0.04b 0.8 ± 0.19b 1.5 ± 0.09ab 1.3 ± 0.16ab 0.8 ± 0.05b 1.9 ± 0.45ab 1.0 ± 0.17b 0.6 ± 0.06b 0.004 n-6 DI4 0.30 ± 0.04 0.47 ± 0.04 0.38 ± 0.02 0.36 ± 0.07 0.44 ± 0.03 0.36 ± 0.04 0.33 ± 0.02 0.41 ± 0.03 0.36 ± 0.07 0.40 ± 0.03 0.25 Mean values within a row with unlike superscript letters were significantly different (P < 0.05) by ANOVA followed by Tukey’s honestly significant difference post hoc test. nd,
746
not detectable levels.
747
1Includes 14:0 and 20:0
748
2Includes 16:1 n-7 and 20:1 n-9
749
3Includes 18:3 n-6 and 20:2 n-6
750
4n-6DI = (20:3 n-6 + 20:4 n-6) / (18:2 n-6 + 20:3 n-6 + 20:4 n-6)
751
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Table 34. Fate of radioactivity from [1-14C] 18:3n-3 incubated in Atlantic salmon hepatocytes in the presence or absence of lipoic acid isolated 752
from fish fed 10 experimental diets containing different levels of EPA and/or DHA for 26 weeks. Values are means ± sem (n = 3).
753
Cellular lipids (nmol) Secreted lipids in media (nmol) CO2 in medium (nmol) ASPF in medium (nmol) Recovery (%)
CONT LA CONT LA CONT LA CONT LA CONT LA
0 % 13.9 ± 0.3 13.5 ± 0.4 2.40 ± 0.12 2.56 ± 0.12 0.021 ± 0.003 0.014 ± 0.002 0.56 ± 0.02 0.56 ± 0.04 80.5 ± 1.0 79.4 ± 2.8 0.5% EPA 12.6 ± 0.3 12.2 ± 0.3 2.60 ± 0.12 2.78 ± 0.08 0.013 ± 0.001 0.008 ± 0.001 0.49 ± 0.03 0.54 ± 0.07 74.8 ± 1.4 73.9 ± 2.0 1.0% EPA 12.1 ± 0.5 12.0 ± 0.4 3.36 ± 0.06 3.40 ± 0.17 0.017 ± 0.002 0.010 ± 0.001 0.56 ± 0.06 0.93 ± 0.24 76.4 ± 2.3 78.0 ± 1.7 2.0% EPA 13.4 ± 1.0 13.7 ± 0.7 2.47 ± 0.27 2.90 ± 0.59 0.016 ± 0.003 0.015 ± 0.003 0.52 ± 0.05 0.63 ± 0.08 78.0 ± 3.5 81.9 ± 0.4 0.5% DHA 12.1 ± 0.9 12.6 ± 0.4 2.87 ± 0.57 2.91 ± 0.70 0.019 ± 0.001 0.013 ± 0.001 0.57 ± 0.05 0.73 ± 0.12 74.1 ± 2.1 77.3 ± 2.2 1.0% DHA 11.2 ± 0.5 10.9 ± 0.5 4.12 ± 0.48 4.06 ± 0.15 0.018 ± 0.001 0.012 ± 0.001 0.62 ± 0.03 0.78 ± 0.06 76.2 ± 1.2 75.1 ± 3.2 2.0% DHA 12.4 ± 1.1 12.0 ± 0.8 3.31 ± 0.42 3.82 ± 0.19 0.021 ± 0.003 0.016 ± 0.003 0.63 ± 0.04 0.87 ± 0.07 78.0 ± 5.5 79.4 ± 4.4 0.5% EPA+DHA 13.1 ± 1.3 13.4 ± 0.2 2.19 ± 0.30 2.14 ± 0.30 0.038 ± 0.023 0.011 ± 0.000 0.51 ± 0.03 0.56 ± 0.09 75.6 ± 4.8 76.7 ± 2.7 1.0% EPA+DHA 11.4 ± 1.1 11.4 ± 0.7 3.52 ± 0.45 3.55 ± 0.53 0.019 ± 0.002 0.015 ± 0.001 0.61 ± 0.03 0.74 ± 0.06 74.1 ± 3.8 74.6 ± 1.0 2.0% EPA+DHA 12.6 ± 0.8 12.0 ± 1.0 3.43 ± 0.55 3.74 ± 0.57 0.017 ± 0.001 0.020 ± 0.004 0.58 ± 0.01 0.80 ± 0.02 79.2 ± 1.7 79.0 ± 2.4
ANOVA
P Diet 0.013 0.0002 0.58 0.02 0.32
P LA 0.69 0.36 0.01 < 0.0001 0.52
P Diet x LA 0.99 0.99 0.47 0.47 0.99
Values are means ± sem (n = 3). ASPF, acid- soluble productsfraction. CONT, control cells; LA, cells supplemented with lipoic acid.
754
Equal amount of cells was seeded for each experimental condition, corresponding to 5.6 ± 0.25 mg protein per flask (mean ± SEM).
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28
Table 45. Percentage distribution between phospholipids (PL), mono- and diacylglycerol (MDG), triglycerides (TAG), and cholesterol esters 757
(CE) produced from [1-14C] 18:3n-3 in Atlantic salmon hepatocytes in the presence or absence of lipoic acid from fish fed different levels of EPA 758
and/or DHA for 26 weeks prior to the experiment (means ± sem; n = 3) 759
PL (%) MDG (%) TAG (%) CE (%)
CONRTROL LA CONRTROL LA CONRTROL LA CONRTROL LA
0 % 90.9 ± 0.9 86.8 ± 0.7 1.3 ± 0.6 1.5 ± 0.4 7.5 ± 0.9 11.5 ± 0.5 0.3 ± 0.0 0.1 ± 0.1 0.5% EPA 89.2 ± 1.8 80.2 ± 1.9 0.5 ± 0.3 1.9 ± 0.2 10.2 ± 1.9 17.7 ± 1.9 nd 0.2 ± 0.2 1.0% EPA 87.7 ± 0.5 75.4 ± 2.4 2.0 ± 0.3 3.0 ± 0.8 10.2 ± 0.3 21.2 ± 1.6 0.1 ± 0.1 0.4 ± 0.1 2.0% EPA 74.8 ± 4.1 63.7 ± 1.5 1.4 ± 0.3 3.2 ± 1.1 23.5 ± 3.7 32.2 ± 1.0 0.4 ± 0.1 0.9 ± 0.2 0.5% DHA 89.8 ± 2.4 81.3 ± 1.5 1.3 ± 0.1 2.3 ± 0.1 8.8 ± 2.3 15.8 ± 1.4 0.1 ± 0.1 0.6 ± 0.0 1.0% DHA 88.6 ± 2.5 75.3 ± 0.6 2.7 ± 0.7 3.5 ± 0.6 8.4 ± 1.8 20.6 ± 1.1 0.2 ± 0.1 0.6 ± 0.3 2.0% DHA 84.3 ± 0.9 74.7 ± 2.1 1.8 ± 0.2 2.9 ± 0.3 13.6 ± 0.6 21.7 ± 1.8 0.4 ± 0.2 0.8 ± 0.1 0.5% EPA+DHA 84.0 ± 6.3 77.1 ± 6.7 1.6 ± 0.9 2.0 ± 1.2 14.2 ± 5.3 20.3 ± 5.6 0.1 ± 0.1 0.6 ± 0.0 1.0% EPA+DHA 87.5 ± 2.0 77.0 ± 2.8 2.4 ± 1.0 2.7 ± 0.8 10.0 ± 2.6 19.6 ± 2.1 0.1 ± 0.0 0.8 ± 0.2 2.0% EPA+DHA 80.3 ± 2.8 69.9 ± 1.9 2.7 ± 1.4 3.6 ± 1.2 16.7 ± 2.8 25.6 ± 3.4 0.3 ± 0.1 0.9 ± 0.3
ANOVA
P Diet < 0.0001 0.14 < 0.0001
P LA < 0.0001 0.009 < 0.0001
P Diet x LA 0.91 0.99 0.91
Total radioactivity recovered in the different lipid classes was set to 100% for each dietary group (PL+ MDG + TAG + CE = 100%). The different P-values are significance levels
760
from two-way ANOVA. nd = non detectable amounts; CONT = control cells; LA = cells supplemented with lipoic acid.
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Table 56. Percentage of substrate added recovered in PUFA from the phospholipid fraction of hepatocytes incubated with [1-14C] 18:3n-3 in 763
lipoic acid free or supplemented media (means ± sem; n = 3). The fish had been fed diets containing different levels of EPA and/or DHA for 26 764
weeks prior to the experiment.
765
18:3n-3 18:4n-3 20:3n-3 20:4n-3 20:5n-3 22:5n-3 22:6n-3
CONT LA CONT LA CONT LA CONT LA CONT LA CONT LA CONT LA
0% 36.4 ± 0.9 30.1 ± 0.7 1.0 ± 0.13 1.4 ± 0.12 0.5 ± 0.03 2.6 ± 0.4 5.4 ± 0.33 4.4 ± 0.29 10.2 ± 0.5 6.8 ± 0.4 1.0 ± 0.08 1.1 ± 0.20 5.3 ± 0.5 8.2 ± 1.1 0.5% EPA 28.6 ± 1.2 21.9 ± 2.0 0.8 ± 0.09 0.9 ± 0.23 0.6 ± 0.28 3.7 ± 0.71 3.5 ± 0.62 2.5 ± 0.32 12.7 ± 0.5 7.4 ± 0.7 0.9 ± 0.11 0.8 ± 0.18 6.0 ± 0.7 8.4 ± 0.6 1.0% EPA 26.6 ± 0.9 22.1 ± 0.1 0.6 ± 0.19 0.5 ± 0.17 1.8 ± 0.47 4.7 ± 0.59 2.3 ± 0.83 1.5 ± 0.55 10.8 ± 2.0 6.4 ± 1.7 0.6 ± 0.11 0.4 ± 0.13 7.3 ± 0.1 7.0 ± 0.6 2.0% EPA 23.8 ± 1.4 19.6 ± 1.4 0.4 ± 0.10 0.4 ± 0.06 3.8 ± 1.11 8.3 ± 1.42 1.7 ± 0.30 1.0 ± 0.14 8.0 ± 1.3 3.9 ± 0.6 0.5 ± 0.06 0.2 ± 0.04 8.0 ± 0.1 6.4 ± 1.0 0.5% DHA 27.6 ± 1.1 24.3 ± 0.8 0.7 ± 0.27 0.7 ± 0.27 1.2 ± 0.33 4.5 ± 0.71 3.1 ± 0.96 2.0 ± 0.50 10.4 ± 2.4 6.4 ± 1.3 1.0 ± 0.27 0.7 ± 0.18 7.0 ± 0.4 8.3 ± 1.4 1.0% DHA 26.2 ± 1.0 22.0 ± 1.0 0.5 ± 0.03 0.4 ± 0.09 2.1 ± 0.24 2.6 ± 1.08 1.4 ± 0.12 1.0 ± 0.09 9.4 ± 1.4 7.3 ± 1.6 0.3 ± 0.05 0.1 ± 0.09 6.9 ± 0.7 5.3 ± 0.6 2.0% DHA 26.6 ± 3.0 22.2 ± 1.8 0.4 ± 0.05 0.3 ± 0.04 5.0 ± 0.95 8.5 ± 1.84 1.3 ± 0.30 0.7 ± 0.06 5.7 ± 0.9 3.3 ± 0.5 0.2 ± 0.19 0.1 ± 0.07 8.7 ± 0.5 5.7 ± 0.4 0.5% E+D 27.3 ± 1.5 22.2 ± 2.9 1.1 ± 0.42 1.3 ± 0.31 0.7 ± 0.33 3.7 ± 1.42 4.1 ± 0.62 3.1 ± 0.40 11.0 ± 0.4 6.6 ± 0.5 1.1 ± 0.20 0.7 ± 0.15 6.0 ± 1.2 10.0 ± 1.9 1.0% E+D 27.1 ± 0.0 21.5 ± 1.3 0.5 ± 0.00 0.4 ± 0.09 2.3 ± 0.00 5.9 ± 1.62 2.0 ± 0.00 1.1 ± 0.04 8.4 ± 0.0 4.6 ± 0.9 0.7 ± 0.00 0.4 ± 0.11 7.9 ± 0.0 6.9 ± 1.0 2.0% E+D 25.1 ± 2.7 20.6 ± 2.6 0.4 ± 0.06 0.3 ± 0.04 4.7 ± 0.38 8.5 ± 0.35 1.5 ± 0.26 0.8 ± 0.13 6.9 ± 1.1 3.5 ± 0.7 0.3 ± 0.10 0.1 ± 0.02 8.8 ± 1.3 5.6 ± 0.7
ANOVA P Diet < 0.0001 < 0.0001 < 0.0001 < 0.0001 0.0003 < 0.0001 0.75
P LA < 0.0001 0.81 < 0.0001 0.0002 < 0.0001 0.01 0.97
P Diet x LA 0.99 0.95 0.69 0.99 0.96 0.85 0.0016
Minor amounts of radioactivity (0.48 ± 0.04%; mean ± sem) were recovered in two non-identified peaks. CONT, control cells; LA, cells supplemented with lipoic acid.
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Table 67. Percentage of substrate added recovered in PUFA from the neutral lipid fraction of hepatocytes incubated with [1-14C] 18:3n-3 in lipoic 768
acid free or supplemented media (means ± sem; n = 3). The fish had been fed diets containing different levels of EPA and/or DHA for 26 weeks 769
prior to the experiment.
770
18:3n-3 18:4n-3 20:3n-3 20:4n-3 20:5n-3 22:5n-3 22:6n-3
CONT LA CONT LA CONT LA CONT LA CONT LA CONT LA CONT LA
0% 3.6 ± 0.5 4.5 ± 0.5 0.5 ± 0.03 0.3 ± 0.07 0.1 ± 0.06 1.6 ± 0.1 0.14 ± 0.14 0.26 ± 0.02 0.58 ± 0.11 0.53 ± 0.20 nd nd 0.77 ± 0.18 1.10 ± 0.11 0.5% EPA 3.1 ± 0.8 5.6 ± 0.6 0.7 ± 0.32 0.8 ± 0.16 0.5 ± 0.24 2.8 ± 0.50 0.11 ± 0.11 0.14 ± 0.09 0.86 ± 0.21 0.49 ± 0.04 0.10 ± 0.06 0.02 ± 0.02 0.82 ± 0.05 1.46 ± 0.14 1.0% EPA 3.5 ± 0.2 6.6 ± 0.8 0.4 ± 0.08 0.5 ± 0.08 1.0 ± 0.10 3.8 ± 0.32 0.06 ± 0.06 0.04 ± 0.04 0.70 ± 0.14 0.69 ± 0.10 nd nd 1.15 ± 0.15 2.25 ± 0.22 2.0% EPA 8.6 ± 2.0 11.4 ± 0.6 0.5 ± 0.05 0.6 ± 0.03 3.5 ± 1.20 7.8 ± 1.23 0.10 ± 0.05 0.21 ± 0.16 0.83 ± 0.06 0.57 ± 0.13 0.03 ± 0.03 nd 2.08 ± 0.41 2.76 ± 0.13 0.5% DHA 3.3 ± 0.9 5.5 ± 0.6 0.4 ± 0.12 0.4 ± 0.09 0.4 ± 0.22 2.6 ± 0.44 0.06 ± 0.06 0.14 ± 0.10 0.43 ± 0.17 0.51 ± 0.17 nd nd 0.86 ± 0.20 1.73 ± 0.19 1.0% DHA 3.5 ± 0.6 6.2 ± 0.7 0.3 ± 0.13 0.5 ± 0.10 0.7 ± 0.45 2.6 ± 0.28 0.04 ± 0.04 0.04 ± 0.04 0.61 ± 0.15 1.16 ± 0.23 nd nd 1.14 ± 0.44 2.40 ± 0.17 2.0% DHA 4.8 ± 0.2 6.4 ± 0.3 0.3 ± 0.06 0.3 ± 0.07 2.2 ± 0.13 5.1 ± 0.39 nd 0.03 ± 0.03 0.31 ± 0.03 0.34 ± 0.18 nd nd 1.55 ± 0.18 1.95 ± 0.44 0.5% E+D 5.0 ± 2.2 7.5 ± 2.0 1.4 ± 0.93 1.0 ± 0.67 1.0 ± 0.31 2.9 ± 0.24 0.48 ± 0.48 0.30 ± 0.30 1.05 ± 0.58 0.50 ± 0.27 nd nd 1.13 ± 0.28 1.61 ± 0.34 1.0% E+D 3.5 ± 0.0 5.9 ± 0.8 0.3 ± 0.00 0.5 ± 0.11 1.2 ± 0.00 3.9 ± 0.77 nd nd 0.56 ± 0.00 0.36 ± 0.18 nd nd 1.15 ± 0.00 1.64 ± 0.34 2.0% E+D 6.3 ± 0.7 8.1 ± 0.1 0.4 ± 0.08 0.3 ± 0.09 2.4 ± 0.35 5.8 ± 0.22 0.08 ± 0.08 0.17 ± 0.10 0.75 ± 0.08 0.40 ± 0.11 0.04 ± 0.04 0.07 ± 0.07 1.66 ± 0.17 2.28 ± 0.06
ANOVA P Diet < 0.0001 0.08 < 0.0001 0.20 < 0.0001
P LA < 0.0001 0.91 < 0.0001 0.20 < 0.0001
P Diet x
LA 0.99 0.99 0.34 0.31 0.65
Minor amounts of radioactivity (0.12 ± 0.02%; mean ± sem) were recovered in two non-identified peaks. nd = non detectable amounts; CONT = control cells; LA = cells
771
supplemented with lipoic acid.
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Table 78. Spearman’s correlation coefficients between cellular EPA or cellular DHA and 18:3n-3 and its FA products in control hepatocytes and 774
hepatocytes supplemented with lipoic acid.
775
Cellular EPA Cellular DHA
CONT LA CONT LA
18:3n-3 -0.325 (0.091) -0.443 (0.014) -0.519 (0.005) -0.392 (0.032) 18:4n-3 -0.257 (0.187) -0.310 (0.096) -0.663 (0.0001) -0.754 (< 0.0001) 20:3n-3 0.393 (0.039) 0.454 (0.012) 0.883 (< 0.0001) 0.582 (0.0007) 20:4n-3 -0.284 (0.144) -0.322 (0.082) -0.763 (< 0.0001) -0.820 (< 0.0001) 20:5n-3 -0.085 (0.668) -0.291 (0.119) -0.6842 (< 0.0001) -0.589 (0.0006) 22:5n-3 -0.275 (0.157) -0.306 (0.100) -0.775 (< 0.0001) -0.854 (<0.0001) 22:6n-3 0.390 (0.040) -0.201 (0.287) 0.767 (< 0.0001)
CONT, control cells; LA, cells supplemented with lipoic acid.
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-2000 -1500 -1000 -500 0
ΔFA in PL betweenControl and cells(pmol)
-500 0 500 1000 1500
0 % 0.5% EPA 1.0% EPA 2.0% EPA 0.5% DHA 1.0% DHA 2.0% DHA EPA+DHA EPA+DHA EPA+DHA
ΔFA in NL betweenControl and LA exposedcells(pmol)
B
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-3000 -2500 -2000 -1500 -1000 -500
ΔFA in PL between0% and inc controlcells(pm
-200 0 200 400 600 800 1000 1200 1400 1600
.5% EPA .0% EPA .0% EPA .5% DHA .0% DHA .0% DHA PA+DHA PA+DHA PA+DHA
ΔFA in NL between 0% and increasing n-3 in control cells (pmol)
B
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0 500 1000 1500 2000 2500 3000
PL+NL [1-14C] 22:6n-3 (pmol) CONT LA
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0% Diet 0.5% Diets 1.0% Diets 2.0% Diets
Relative expression
srebp1
0% Diet 0.5% Diets 1.0% Diets 2.0% Diets