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Supporting informtion Multiple Univrite Dt Anlysis Revels the Inulin Effects on the High-ft-diet Induced Metolic Altertions in Rt Myocrdium nd Testicles in the Pre-oesity Stte Yixun Dun #, Ynpeng An #, Ning Li #, Bifeng Liu, Yuln Wng nd Huiru Tng * Britton Chnce Center for Biomedicl Photonics t Wuhn Ntionl Lortory for Optoelectronics-Huei Bioinformtics nd Moleculr Imging Key Lortory, Systems Biology Theme, Deprtment of Biomedicl Engineering, College of Life Science nd Technology, Huzhong University of Science nd Technology, Wuhn,, P. R. Chin Key Lortory of Mgnetic Resonnce in Biologicl Systems, Stte Key Lortory of Mgnetic Resonnce nd Atomic nd Moleculr Physics, Centre for Biospectroscopy nd Metonomics, Wuhn Institute of Physics nd Mthemtics, the Chinese Acdemy of Sciences, Wuhn, 9, P. R. Chin University of Chinese Acdemy of Sciences, Beijing 9, P. R. Chin *To whom ll correspondence should e ddressed. Huiru Tng: e-mil, Huiru.tng@wipm.c.cn; tele,+--9; fx, +--999.

Figure legends: Figure S. Schemtic illustrtions for niml experiments. Figure S. Chnges of (A) ody weight, (B) verge dily energy intke, (C) ody-weight-gin per unit of energy intkes (mg/kcl) for rts fed with norml diet (ND), control diet (CD), inulin-contining control diet (ICD), high-ft diet (HFD) nd inulin-contining HFD (IHFD). Dotted lines indicted the diet-switch time. Ech vlue is men ± S.D. CD vs. HFD p<.5; HFD vs. IHFD p<.5; c CD vs. IHFD p<.5. Figure S. Hert model evlution sed upon pirwise comprison of (A) CD vs. HFD, (B) HFD vs. IHFD, nd (C) CD vs. IHFD. From left to right re PCA scores plots, PLS-DA scores plots, rndom permuttion tests, nd OPLS-DA scores plots. Figure S. Testicle model evlution sed upon pirwise comprison of (A) CD vs. HFD, (B) HFD vs. IHFD, nd (C) CD vs. IHFD. From left to right re PCA scores plots, PLS-DA scores plots, rndom permuttion tests, nd OPLS-DA scores plots. Figure S5. OPLS-DA loding plots showing the discrimintion etween hert hydrophilic extrcts of (A) CD vs. HFD, (B) HFD vs. IHFD, nd (C) CD vs. IHFD. (Forced implementtion) Mgnifiction fctors of the regions δ 5.-9. reltive to δ.5-.. Figure S. OPLS-DA loding plots showing the discrimintion etween testicle hydrophilic extrcts of (A) CD vs. HFD, (B) HFD vs. IHFD, nd (C) CD vs. IHFD. (Forced implementtion) Mgnifiction fctors of the regions δ 5.-9. reltive to δ.5-.. Figure S. Results from the Student s t-test (if criteri met) or Kruskl-Wllis test of metolites in myocrdium from rts to vlidte the results from the multiple univrite dt nlysis. Verticl xis: pek re per mg tissue. CD vs. HFD, p<.5; CD vs. IHFD, p<.5; c HFD vs. IHFD, p<.5. Keys: Tu, turine; Lc, lctte; Glu, glutmte; GPC, glycerophosphocholine; AMP, denosine monophosphte; Lys, lysine; -HB, -hydroxyutyrte; HTu, hypoturine; HpX, hypoxnthine; NAD +, nicotinmide denine dinucleotide; SI, scyllo-inositol. Figure S. Results from the Student s t-test (if criteri met) or Kruskl-Wllis test of metolites in testicle tissues from rts to vlidte the results from the multiple univrite dt nlysis. Verticl xis: pek re per mg tissue. CD vs. HFD, p<.5; CD vs. IHFD, p<.5; c HFD vs. IHFD, p<.5. Keys: Glu, glutmte; Al, lnine; Thr, threonine; HpX, hypoxnthine; -HB, -hydroxyutyrte; Xn, xnthine; Succ, succinte; His, histidine; SI, scyllo-inositol.

Tle S. Formul for control (CD) nd high-ft diet (HFD) CD HFD Ingredient g kcl g kcl Csein, Mesh L-Cystine Com Strch 5. 9 Mltodextrin 5 Sucrose 5. 9 Cellulose, BW 5 5 Soyen Oil 5 5 5 5 Lrd.5 59 Minerl Mix S Diclcium Phosphte Clcium Cronte 5.5 5.5 Potssium Citrte, H O.5.5 Vitmin Mix V Choline Bitrtrte Totl 55 5 5. 5

Tle S. Results from the Student s t-test (if criteri met) or Kruskl-Wllis test for some metolites in myocrdium tissues from rts fed with control diet (CD), high-ft diet (HFD) nd inulin-contining high-ft diet (IHFD) Metolites Signl re per mg tissue p-vlues from Student s t-test CD HFD IHFD CD vs.hfd HFD vs. IHFD CD vs.ihfd Glutmte 5.±. 5.9±. 5.±..9 - -. - Lysine.±..9±..±.. - -. - Turine.±..5±. 9.5±.5 - -. - Hypoturine.5±..±.5.±.5 5. -5.9 -.5 - -Hydroxyutyrte.5±..±..9±.. - - - Lctte.±.5.±..±..5 - - - scyllo-inositol.±..±..±. - 9.9 -.5 - GPC.±..±..55±..5 -. - - AMP.±..±.5.±.. - -. - Hypoxnthine.±.5.±..±..9 - -. - Inosine.9±..±..±.. - - - Gunosine.9±..9±..±. -.99 - - NAD +.9±..±..±. 5. - -. - NMR signl integrls of metolite re linerly correlted to its concentrtion. Vlues here re expressed s pek-re per mg tissue (men ± SD). only those with p<.5 were tulted here. GPC, glycerophosphocholine; AMP, denosine monophosphte; NAD +, nicotinmide denine dinucleotide.

Tle S. Results from the Student s t-test (if criteri met) or Kruskl-Wllis test for some metolites in testicle tissues from rts fed with control diet (CD), high-ft diet (HFD) nd inulin-contining high-ft diet (IHFD) Metolites Signl re per mg tissue p-vlues from Student s t-test CD HFD IHFD CD vs.hfd HFD vs. IHFD CD vs. IHFD Alnine.±..±..9±. 5. -.9 -.9-5 Glutmte 5.5±.5 5.±.9 5.±.. - -. - Threonine.±.5.±.9.9±.9 -. -. - Histidine.5±..5±..±.5 - -.9 - -Hydroxyutyrte.±. 5.5±..9±..9 - - - Succinte.99±..±..±.. - - - Acette.9±..9±..±. -. -. - scyllo-inositol.9±..±..±. -.9 -. -9 Betine.5±..±.5.±.. -. - - Ure.±..9±..±..9 -. -.9 - Urcil.9±..95±.9.±. - -.9 - Xnthine.9±..±..±. - -.9 - Hypoxnthine 5.±.5.±.55.±.. - - 5.9 - Inosine.5±..±.5.±. 9.9 - - - NMR signl res of metolite re linerly correlted to its concentrtion. Vlues here re expressed s pek-re per mg tissue (men ± SD). only those with p<.5 were tulted here.

Figure S. IHFD group w 5w w HFD group CD group w d 5w w Norml diet (Zhongnn Hospitl, Wuhn, Chin) Control diet (CD) High-ft diet (HFD) Inulin-contining control diet (ICD) Inulin-contining high-ft diet (IHFD)

Figure S. A) Body weight(g) B) Energy Intke (kcl). 5.. 55. 5. 5.. 5.. 5. 9 ND CD HFD ICD IHFD c c c c c c c c c c c c - - - 5 9 c c c c c C) Body-weight-gin ginst energy-intke (mg/kcl) 5 5 - - - - - - - 5 9 c c c c c - - - 5 9 c c Time (w)

Figure S. R X=.5, Q =... Q =.5, p=. - CD IHFD C) 5.. O -5 - -.. - - - -. - - - - R X=., Q =-.......5....9... - - - P Q =., p=5. - HFD IHFD B) 5.. O -5 -. - -. - - - -. - - - - - R X=., Q =.......5....9... - - - P Q =., p=9.5 - CD HFD A) 5.. O -5 -.. - - - -. - - - -......5....9. - - P

Figure S. R X=., Q =.9.. Q =., p=. CD IHFD C)... O - -. - - -. -5 5 - - R X=., Q =-......... - - P Q =.9, p=. - HFD IHFD. B). O. - - -. - - -. -5 5 - - R X=.9, Q =.5......... - P Q =., p=.9 CD HFD A) - -... O - - - -. -5 5 - -...... - - P

Figure S5..5 x.5 C) x IHFD -.5 - - -.5 9.5 9.5.5.5 5.5 - -.5.5.5.5 CD.5 x 5 5 B) x IHFD - - - - 9.5 9.5.5.5 5.5 - -.5.5.5.5 HFD.5 5 x 5 A) x.5 HFD.5 -.5 - -5 9.5 9.5.5.5 5.5 -.5 -.5.5.5.5 CD.5

Figure S. x 5 C) x IHFD - - - - 9.5 9.5.5.5 5.5 - -.5.5.5.5 CD.5 x 5 B) x IHFD - - - - - - 9.5 9.5.5.5 5.5 - -.5.5.5.5 HFD.5.5 x A) x.5 HFD.5 -.5 -.5 - - 9.5 9.5.5.5 5.5 -.5 -.5.5.5.5 CD.5

Figure S. Reltive concentrtion (to weight of tissue) 5 5 5 5..... Tu Lys Lc -HB 5..5.... c CD HFD IHFD Glu Inosine GPC AMP c c HTu HpX Gunosine NAD+ SI c

Figure S. Reltive concentrtion (to weight of tissue) 5 5 Glu 5 5 5 c CD HFD IHFD c c Betine Al Thr Inosine c........ His c SI c HpX -HB Xn Acette Succ Ure Urcil