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1 Supplementry Figure S1. Ft-1 mice exhibit reduced diposity when fed n HFHS diet. WT nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. A: Representtive photogrphs for side-by-side comprison of mle WT nd trnsgenic mice fed the HFHS diet for 18 weeks. B: Epididyml ft mss in WT nd ft-1 mice fed HFHS diet (n=8 per group). C: Body composition nlysis (ft mss nd len mss) of WT nd ft-1 mice before nd fter 18 weeks on the HFHS diet using dul-energy X-ry bsorptiometry. n= 10 to 12 for control nd HFHS groups. *P<0.05; **P<0.01 (Mnn-Whitney test test). Similr results were obtined in five independent experiments. D: UCP1 mrna expression in interscpulr brown dipose tissue. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test) Americn Dibetes Assocition. Published online t

2 Supplementry Figure S2. F4/80 immunohistochemistry lbeling in dipose tissue from wild type nd ft-1 mice fed control or high ft high sucrose diet. Box nd whisker digrm showing lbelling index (rtio between F4/80 re nd tissue re) for the different groups. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test). WT CTL WT HFHS Ft-1 CTL Ft-1 HFHS F4/80 re / tissue re rtio b WT CTL WT HFHS Ft-1 CTL Ft-1 HFHS 2018 Americn Dibetes Assocition. Published online t

3 Supplementry Figure S3. Ft-1 mice disply resistnce to HFHS-induced hyperglycemi nd insulin resistnce. WT nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. A: The mice were fsted for 12h for glycemi nd insulinemi determintion. B: GLUT1 mrna expression in skeletl muscle C: Dose-response histogrms of insulin-stimulted glucose trnsport in isolted gstrocnemius muscles from WT nd ft-1 mice fed n HFHS diet fter overnight food deprivtion. n=4 (A nd B); n=8 (C); n=6 (D). Dt re expressed s the men ± SEM. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test). *P<0.05; **P<0.01 (Mnn-Whitney test). Similr results were obtined in five independent experiments Americn Dibetes Assocition. Published online t

4 Supplementry Figure S4. Muscle lipidomic nlysis. Wild-type nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. Muscle mjor ftty cid composition, totl omeg 6, totl omeg 3 nd omeg6/omeg3 rtios re indicted for WT nd ft-1 mice fed with control or HFHS diets (n=6). The results re expressed s the men ± SEM. Dt with different superscript letters re significntly different t P<0.05, ccording to the Kruskl-Wllis test nd the Dunn's multiple comprisons test Americn Dibetes Assocition. Published online t

5 Supplementry Figure S5. Omeg 3 ftty cid enrichment protects ft-1 mice ginst liver HFHSinduced lipogenesis. Wild-type nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. A: Liver totl lipid levels in mice fed either control or HFHS diet. B: Liver mjor ftty cid composition, totl omeg 6 nd totl omeg 3 re indicted for WT nd ft-1 mice fed with control or HFHS diets (n=8). C: Heptic FATCD36 nd PPAR mrna levels were mesured by qrt-pcr. All mrna levels were normlized reltive to -ctin (n=10). The results re expressed s the men ± SEM. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test). FATCD36: Ftty cid trnsporter cluster of differentition 36; PPAR peroxisome prolifertor-ctivted receptor gmm. Similr results were obtined in five independent experiments Americn Dibetes Assocition. Published online t

6 Supplementry Figure S6. Food intke in ft-1 nd WT mice fed the control or HFHS diet. 24h food intke (recorded every 3 dys over 3 weeks) is expressed in wild-type nd ft-1 trnsgenic mice fed control diet (CTL) nd high ft/high sucrose (HFHS) diet. n= 7 for ech group. The results re expressed s the men ± SEM. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test) Americn Dibetes Assocition. Published online t

7 Supplementry Figure S7. DHA supplementtion hs no direct effect on permebility of cultured Cco2-TC7 monolyers. A: Effect of DHA on FITC-Dextrn permebility. B: ZO-1 mrna expression. Cells were treted with 100 M of DHA for 24 h nd BSA tretment served s control. Results re presented s men ± SEM. FITC-Dextrn: Fluorescein Isothiocynte-dextrn; ZO-1: Zonul Occludens 1; DHA: docoshexenoic cid; BSA: bovine serum lbumin Americn Dibetes Assocition. Published online t

8 Supplementry Figure S8. Ft-1 trnsgene expression long the intestinl trct. The expression of the ft-1 trnsgene ws ssessed in the distinct segments of the intestinl trct (duodenum, jejunum, ileum, cecum nd colon) in wild-type nd trnsgenic nimls. WT Ft-1 Duodenum Jejunum Ileum Cecum Colon 2018 Americn Dibetes Assocition. Published online t

9 Supplementry Figure S9. Food intke in WT, Ft-1 nd Ft-1-to-WT mice fed the HFHS diet. The levels of 24h food intke (recorded every 2 dys for 2 weeks period) is expressed in the wild-type, Ft-1 nd Ft-1 to WT mice fed high ft/high sucrose (HFHS) diet. N= 6 for ech group. The results re expressed s the men ± SEM. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test). WT Ft-1 Ft-1 to WT 2018 Americn Dibetes Assocition. Published online t

10 Supplementry Figure S10. Antimicrobil peptides expression in the jejunum nd ileum of control nd high ft high sucrose-fed WT nd ft-1 mice. Expression of Regenerting islet-derived 3-gmm (RegIIIγ, encoded by Reg3g); Phospholipse A2 group II (encoded by Pl2g2) nd α-defensins (encoded by Def), mesured in the jejunum nd ileum of control diet-fed mice (CTL) nd high ft high sucrosefed mice (HFHS). N= 6 for ech group. The results re expressed s the men ± SEM. Groups with different superscript letters re significntly different (P<0.05, ccording to Kruskl-Wllis test nd the Dunn's multiple comprisons test) Americn Dibetes Assocition. Published online t

11 Supplementry Tble 1. The sequences of primers used in Reltime PCR ssys. GENES Forwrd Primer Reverse Primer FAT/CD36 5 -AATTAGTAGAACCGGGCCAC-3 5 -CCAACTCCCAGGTACAATCA-3 PPAR 5 -ATCTTAACTGCCGGATCCAC-3 5 -AGGCACTTCTGAAACCGACA-3 ZO-1 5 -GACCTTGAGCAGCCGTCATA-3 5 -CCGTAGGCGATGGTCATAGTT-3 UCP-1 5 -GCTTAATGACTGGAGGTGTGG-3 5 -CTCTGGGCTTGCATTCTGAC-3 GLUT-1 5 -GTGACGATCTGAGCTACGGG-3 5 -TCACCTTCTTGCTGCTGGG-3 Def 5 -GGTGATCATCAGACCCCAGCATCAGT-3 5 -AAGAGACTAAAACTGAGGAGCAGC-3 Reg3g 5 -TTCCTGTCCTCCATGATCAAA-3 5 -CATCCACCTCTGTTGGGTTC-3 Pl2g2 5 -AGGATTCCCCCAAGGATGCCAC-3 5 -CAGCCGTTTCTGACAGGAGTTCTGG-3 Actin 5 -ATGCTCCCCGGGCTGTAT-3 5 -CATAGGAGTCCTTCTGACCCATTC-3 Supplementry Tble 2. Anlyses of the composition of ftty cids in Diets. Molr % Ftty cid D12450B (CTL) D12451 (HFHS) 14: : : : SFA : : : MUFA :2N :3N :2N PUFA n-6/n Abbrevitions: SFA, short-chin ftty cids; MUFA, monounsturted ftty cids; PUFA, polyunsturted ftty cids Americn Dibetes Assocition. Published online t

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