* * * * * liver kidney ileum. Supplementary Fig.S1
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1 Supplementry Fig.S1 liver kidney ileum Fig.S1. Orlly delivered Fexrmine is intestinlly-restricted Mice received vehicle or Fexrmine (100mg/kg) vi per os (PO) or intrperitonel (IP) injection for 5 dys (n=3/group). () Expression of the FXR trget gene SHP fter PO or IP injection in selected tissues. () Expression of the FXR trget genes fter IP Fexrmine tretment in selected tissues. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
2 Supplementry Fig.S2 d c e Fig.S2. Fexrmine hs no effect in norml mice () Body weight, () cron dioxide production, (c) oxygen consumption, (d) glucose tolernce test, nd (e) core temperture of chow-fed mice treted with vehicle or fexrmine (100mg/kg) vi PO for 5 weeks. n=4/group. Dt represent the men ± SD. Sttisticl nlysis s performed with the Student s t test. p<0.05, p<0.01
3 Supplementry Fig.S3 c Fig.S3. Fexrmine dose dependently improves glucose homeostsis. () Fsting glucose levels in 14 week HFD fed mice treted with vehicle or fexrmine (100mg/kg/dy os for 5 week). () Body weights of 14 week HFD fed mice treted with vehicle or fexrmine (10, 50, or 100mg/kg/dy per os) for 5 weeks with HFD. (c) Glucose tolernce test performed on HFD fed mice treted with vehicle or fexrmine (10, 50, or 100mg/kg/dy per os) for 5 weeks with HFD. n=6/group Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
4 Supplementry Fig.S4 ileum colon Vehicle Fexrmine Fig.S4. Fexrmine tretment improves gut helth Histologicl nlysis of ileum nd colon of 14 week HFD fed mice fter vehicle or Fexrmine tretment (100mg/kg/dy os for 5 week with HFD). Scle rs in ileum: 50 μm, scle rs in colon: 100 μm.
5 Supplementry Fig.S5 c d e f g h i Fig.S5. FXR is required for Fexrmine s effects () glucose tolernce test, () insulin sensitivity test, (c) Body weights (d) oxygen consumption, (e) cron dioxide production, (f) core temperture, (g) rown dipose tissue gene expression, (h) liver gene expression, nd (i) FXR trget gene expressions in ileum of 14 week HFD fed FXR-null mice treted with vehicle or fexrmine (100mg/kg) for 5 week with HFD. n=4/group. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
6 Supplementry Fig.S6 Fig.S6. Fexrmine does not ffect the respirtory exchnge rtio. Respirtory exchnge rtios in 14 week HFD fed mice treted with vehicle or fexrmine (100mg/kg/dy os) for 5 week with HFD.
7 Supplementry Fig.S7 pp38 P38 RlA Vehicle Fex(50mg/kg) Fex (100mg/kg) c KEGG pthwy Term Oxidtive phosphoryltion Chemokine signling pthwy Cytokine-cytokine receptor interction Biosynthesis of unsturted ftty cids PPAR signling pthwy p-vlue 8.12E E E E E-03 OXPHOS gene chnges normlized expression d e f iwat gwat BAT Fig.S7. Fexrmine enhnces rown dipose tissue OXPHOS 14 week HFD fed mice were treted with vehicle or fexrmine (100mg/kg/dy os for 5 week on HFD). () Totl nd phosphorylted P38 protein in BAT. () Hetmp of expression chnges OXPHOS genes. (c) KEGG pthwy nlysis of trnscriptionl chnges in BAT. (d) Hetmp of chemokine nd cytokine signling pthwy in BAT. (e) PKA ctivity in BAT. (f) Serum lctte level. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
8 Supplementry Fig.S8 ileum KEGG pthwy Term PPAR signling pthwy Adipocytokine signling pthwy Retinol metolism Drug metolism Archidonic cid metolism p-vlue 1.86E E E E E-03 colon KEGG pthwy Term PPAR signling pthwy Adipocytokine signling pthwy Retinol metolism p-vlue 3.52E E E-02 Fig.S8. Fexrmine induces multiple metolic pthwys Mice fed HFD for 14 weeks were mintined on HFD nd treted with vehicle or fexrmine (100mg/kg/dy os for 5 week). () Kegg pthwy nlysis of trnscriptionl chnges induced in ileum nd colon, s determined y RNA-Seq. () Heptic Cyp71 levels determined y ELISA. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
9 Supplementry Fig.S9 Fig.S9. Fexrmine fils to ctivte TGR5 HEK293 cells were trnsfected with expression vectors for camp-response element luciferse, β-glctosidse nd humn TGR5. 24 hr fter trnsfection, cells were treted with Fexrmine or INT-777 t indicted concentrtion for 5 hr. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
10 Supplementry Fig.S10 % of camp production L A L EC50 300nM RO EC50 70nM c Compound concentrtion, log [M] d e f n.s Fig.S10 Systemic TGR5 ctivtion is required to ffect glucose homeostsis HFD-fed mice were treted with vehicle, the intestinllyrestricted TGR5 lignd L (, L755,100mg/kg EC50 300nM) or the systemic lignd RO (, RO,100mg/kg) vi per os for 14 dys. c, Plsm L755 concentrtion in portl nd til vein fter PO dministrtion. d, Body weight curve. e, Glucose tolernce test. f, Insulin secretion in vivo fter glucose injection. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
11 Supplementry Fig.S11 ileum c d e BAT f g h i j liver soleus
12 Fig.S11. TGR5 is required for suset of Fexrmine e effects HFD-fed TGR5-null mice were treted with vehicle or fexrmine (100mg/kg os dily for 5 week with HFD, n=10). () Ilel FXR trget gene expressions () Serum BA levels (c) Fsting glucose level (left) nd Glucose tolernce test (right) (d) Core temperture (left), Oxygen consumption rte (middle), nd cron dioxide production (right) (e) Gene expressions in BAT (f) Body weight curve (left) nd Body composition y MRI (right) (g) Insulin Tolernce Test (h) Heptic gene expression (i) Heptic TG levels (j) Gene expressions in soleus. n=9/group. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
13 Supplementry Fig.S12 Fig.S12. Fex tretment suppresses inflmmtion in dipose tissues. Mice fed HFD for 14 weeks were mintined on HFD nd treted with vehicle or fexrmine (50mg or 100mg/kg/dy os for 5 week). () Gondl white dipose tissue (gwat) ws sujected to protein expression nlysis y western lotting. () Expression of inflmmtory cytokines in BAT. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
14 Supplementry Fig.S13 Fig.S13. Fexrmine enhnces OXPHOS in iwat Mice fed HFD for 14 weeks were mintined on HFD nd treted with vehicle or fexrmine (100mg/kg/dy os for 5 week). () Gene expressions nd () oxygen consumption rte of SVF from inguinl ft (iwat). Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
15 Supplementry Fig.S14 c clmp m g / k g / m i n IS-GDR Ve h Fex I n s u l i n ( n g / m L ) Ve h Fe x Bsl Clmped Fig.S14. Hyperinsulinemic-euglycemic clmps Mice fed HFD for 14 weeks were mintined on HFD nd treted with vehicle or fexrmine (100mg/kg/dy os for 3 week). Hevier mice (y 2~3 grm) were selected for fexrmine tretment in order to chieve equivlent treted ody weights for the clmp study. () Body weight curve. () Insulinstimulted GDR (IS-GDR). (c) Fsting insulin level. n=14/group. Dt represent the men ± SD. Sttisticl nlysis ws performed with the Student s t test. p<0.05, p<0.01
16 Supplementry Tle.S1 Bile cid composition (%) Vehicle Fexrmine CA TCA CDCA TCDCA LCA GLCA N.D DCA TDCA HDCA T-HDCA 0.99 N.D UDCA T-UDCA lph MCA 0.33 N.D et MCA 0.55 N.D T-et MCA omeg MCA Tle S1. Fexrmine lters the serum ile cid composition Mice fed HFD for 14 weeks were mintined on HFD nd treted with vehicle or fexrmine (100mg/kg/dy per os for 5 week). Serum ile cid composition ws determined y mss spectrometry. N.D not determined
Supplementary Figure S1
Supplementry Figure S Tissue weights (g).... Liver Hert Brin Pncres Len mss (g) 8 6 -% +% 8 6 Len mss Len mss (g) (% ody weight) Len mss (% ody weight) c Tiilis nterior weight (g).6...... Qudriceps weight
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