bead beating method previously described (1). The V4 region of the bacterial 16S rrna gene


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1 1 Supplementl Informtion Gut microbiot nlysis Genomic DNA purifiction n 16S rrna gene mplifiction. Genomic DNA ws isolte from pproximtely 6 mg of cecl content using the repete be beting metho previously escribe (1). The V4 region of the bcteril 16S rrna gene ws mplifie using primers s reporte by Kozich et l (2). PCR ws crrie out uner the following conitions: initil enturtion for 3 min t 94 C, followe by 25 cycles of enturtion for 45 s t 94 C, nneling for 6 s t 5 C n elongtion for 9 s t 72 C, n finl elongtion step for 1 min t 72 C n further trete s reporte previously (3). Sequence nlysis Sequencing ws one t the Genomic Core Fcility of Gothenburg University. The forwr n reverse res from the piren sequencing were joine by exploiting the long overlp between both res using inhouse coes. Ienticl bses in the overlp sequence increse the ssurnce ccurcy of the sequencing n therefore we ssigne the highest possible qulity score for those mtching bses. The FASTXToolkit ws use to filter out lowqulity res n res with qulity Phre score over 2 in t lest 98% of their sequences psse the filter. The sequencing t were then nlyze using the softwre pckge Quntittive Insights Into Microbil Ecology (QIIME), version Sequences were clustere into opertionl txonomic units (OTUs) t 97% ientity threshol using closereference OTU picking pproch with UCLUST (4) ginst the Greengenes reference tbse (5). Representtive sequences for the OTUs were Greengenes reference sequences or cluster sees, n were txonomiclly ssigne using the Greengenes txonomy n the Ribosoml Dtbse Project Clssifier (6). Representtive OTUs were ligne using PyNAST (7) n use to buil
2 2 phylogenetic tree with FstTree (8), which ws use to estimte the βiversity of smples using phylogenetic iversity (9) n weighte unifrc (1). Threeimensionl principl coorintes nlysis plots were visulize using Emperor (11). With this pproch, totl of sequences groupe in 79 OTUs were obtine for the 43 smples sequence, with mein of sequences ssigne to ech smple. To correct for ifferences in sequencing epth, 48 4 sequences were rnomly subsmple for ech smple n use for iversity nlyses. Sequences with very low bunnce (reltive bunnce <.5%) were exclue from the nlysis. LDA Effect Size lgorithm (12) ws use to ientify tx tht iscriminte cecl microbiot profiles ccoring to the coloniztion origin. Bile ci nlysis BAs from 5 µl serum (from portl n cvl veins) were extrcte using protein precipittion with 1 volumes of methnol contining euteriumlbelle internl stnrs (2.5 µm n 5 nm of ech BA stnr for portl n cvl vein respectively) (13). The smples were vortexe for 1 min n then centrifuge t 2g for 1 min. For serum from portl vein, the superntnt ws ilute 1 times in methnol:wter (1:1) n from cvl vein, the superntnt ws evporte n reconstitute in 2 µl of methnol:wter (1:1). BAs from liver n cecum were extrcte fter homogenizing the tissue in 2 ml propylene tubes with 6 zirconium oxie bes (3 mm) (Retsch GmbH, Hn, Germny). Approximtely 5 mg of tissue ws plce in the tube n 5 µl methnol contining internl stnrs were e (2.5µM of ech BA stnr). The tissue ws homogenise using TissueLyser II instrument (Qigen, Hilen, Germny) t 25 Hz for 1 min. The homogente ws then centrifuge t 2 g for 1 min n the superntnt ws ilute 5 times with wter: methnol [1:1] before
3 3 nlysis. BAs from gllbler were extrcte s bove using the whole gllbler n 5 µl of methnol. After homogenistion n centrifugtion, the superntnt ws ilute 1 times with wter:methnol [1:1] before nlysis. Becuse of the high BA concentrtion in gllbler, the internl stnrs (5 nm of ech) were e fter the ilution. BAs were nlyse using ultrhigh performnce liqui chromtogrphy tnem mss spectrometry (UPLCMS/MS). The nlyticl setup consiste of Rheos Allegro qurternry ultrperformnce pump (Flux Instruments, Bsel, Switzerln) n QTRAP 55 mss spectrometer (ABSciex, Concor, Cn). The BAs (5 µl injection) were seprte on Kinetex C18 column (2.1 x 1 mm 1.7 µm prticles) (Phenomenex, Torrnce, CA, USA) kept t 6 C. The mobile phses were 7.5 mm mmonium formte in wter juste to ph 4.5 with formic ci (Aphse) n.1% formic ci in cetonitrile (Bphse). The UPLC grient strte with 1 min isocrtic elution with 25% B. From 1 to 5 min, B ws increse to 35%; from 5 to 14.5 min, B ws increse to 95%; the grient ws then kept t this level for.5 min n then, from 15 to 15.5 min, returne to 25% B. The metho ene with 2.5 min equilibrtion to give totl runtime of 18 min per smple. The flow rte ws 4 µl/min n etection of BAs ws me using scheule MRM in negtive moe. MRM etection winow ws 9 s n trget scn time ws.3 s. Quntifiction ws me ginst clibrtion curves generte from nonlbelle stnrs. Regents Methnol n formic ci were from Merck (Drmstt, Germny). Ammonium formte ws bought from SigmAlrich (Steinheim, Germny) n cetonitrile ws from Rthburn Chemicls Lt (Wlkburn, UK). The BAs turocholic ci (TCA), turoursoeoxycholic ci (TUDCA), turohyoeoxycholic ic (THDCA), turochenoeoxycholic ci (TCDCA) turoeoxycholic ci (TDCA), cholic ci (CA), ursoeoxycholic ci (UDCA), hyoeoxycholic ic (HDCA), chenoeoxycholic ci (CDCA), eoxycholic ci (DCA) n
4 4 lithocholic ci (LCA) were purchse from Sigm Alrich (Stockholm, Sween). Muricholic cis (α, β, n ωmca, turineconjugte n unconjugte) n murieoxycholic ci (MDCA) were purchse from Sterlois (Newport, RI, USA). Deuterte bile cis 4 TCA, 4 UDCA, 4 CDCA n 4 LCAwere purchse from CDN isotopes (Quebec, Cn). Iso DCA ws kin gift from Jn Sjövll n CA7 sulfte ws kin gift from AstrZenec in Mölnl, Sween. In vitro ssy of BA metbolism by humn microbiot Fecl smples from the first humn onor were inoculte (1% w/v) in 5 ml sulfterich nerobic BHI meium (SRANBBHI contining cellobiose (1 g/l), mltose (1 g/l), cysteine (.5 g/l), yest extrct (2.5 g/l), mmonium sulfte (3.31 g/l), hemin (1 mg/l) n BHI (37 g/l)) supplemente with.2% TCA or CA. All chemicls were obtine from Sigm Alrich except BHI which ws purchse from OXOID, UK. The fecl smples were incubte s btch cultures uner strict nerobic conitions mintine in COY chmber for 24 hours t 37 C. The nerobic tmosphere of the COY chmber ws compose of 5% hyrogen, 1% crbon ioxie n 85% nitrogen. Smples (5 µl) were tken for BA nlysis t strt n fter 24 hours' incubtion n were kept in 2 C until the time of nlysis.
5 5 Supplementl References 1. Slonen A, Nikkil J, JlnkTuovinen J, Immonen O, RjilicStojnovic M, Kekkonen RA, Plv A, et l. Comprtive nlysis of fecl DNA extrction methos with phylogenetic microrry: effective recovery of bcteril n rchel DNA using mechnicl cell lysis. J Microbiol Methos 21;81: Kozich JJ, Westcott SL, Bxter NT, Highlner SK, Schloss PD. Development of ulinex sequencing strtegy n curtion pipeline for nlyzing mplicon sequence t on the MiSeq Illumin sequencing pltform. Appl Environ Microbiol 213;79: De Ver F, KovtchevDtchry P, Gonclves D, Viner J, Zitoun C, Duchmpt A, Bckhe F, et l. Microbiotgenerte metbolites promote metbolic benefits vi gutbrin neurl circuits. Cell 214;156: Egr RC. Serch n clustering orers of mgnitue fster thn BLAST. Bioinformtics 21;26: DeSntis TZ, Hugenholtz P, Lrsen N, Rojs M, Broie EL, Keller K, Huber T, et l. Greengenes, chimerchecke 16S rrna gene tbse n workbench comptible with ARB. Appl Environ Microbiol 26;72: Wng Q, Grrity GM, Tieje JM, Cole JR. Nive Byesin clssifier for rpi ssignment of rrna sequences into the new bcteril txonomy. Appl Environ Microbiol 27;73: Cporso JG, Kuczynski J, Stombugh J, Bittinger K, Bushmn FD, Costello EK, Fierer N, et l. QIIME llows nlysis of highthroughput community sequencing t. Nt Methos 21;7: Price MN, Dehl PS, Arkin AP. FstTree 2 Approximtely MximumLikelihoo Trees for Lrge Alignments. PLoS One 21;5:e Fith DP. Conservtion Evlution n Phylogenetic Diversity. Biologicl Conservtion 1992;61: Lozupone C, Knight R. UniFrc: new phylogenetic metho for compring microbil communities. Appl Environ Microbiol 25;71: VázquezBez Y, Pirrung M, Gonzlez A, Knight R. EMPeror: tool for visulizing highthroughput microbil community t. GigScience 213;2: Segt N, Izr J, Wlron L, Gevers D, Miropolsky L, Grrett WS, Huttenhower C. Metgenomic biomrker iscovery n explntion. Genome Biol 211;12:R Tremroli V, Krlsson F, Werling M, Sthlmn M, KovtchevDtchry P, Olbers T, Fnriks L, et l. RouxenY Gstric Bypss n Verticl Bne Gstroplsty Inuce LongTerm Chnges on the Humn Gut Microbiome Contributing to Ft Mss Regultion. Cell Metb 215;22:
6 A Frction 1. D CONVH2 2 w 15 w Verrucomicrobiles TenericutesML615J28 MollicutesRF39 MollicutesAneroplsmtles GmmproteobcteriEnterobcteriles EpsilonproteobcteriCmpylobcterles DeltproteobcteriDesulfovibrionles BetproteobcteriBurkholeriles AlphproteobcteriRF32 Erysipelotrichles Clostriiles Lctobcillles CynobcteriYS2 Bcteroiles Coriobcteriles Bifiobcteriles B Cecum BAs (nmol) w 15 w TαMCA TβMCA TωMCA TCA αmca βmca ωmca CA HDCA DCA C isodca in cecum (nmol) w 15 w D CA 7sulfte in cecum (nmol) w 15 w Supplementl Figure S1. Gut microbiot n BA composition in cecum of mice colonize with secon humn onor. (A) Reltive bunnce of orers in cecl bcteri from CONVH2 mice colonize for 2 weeks (2w) or 15 weeks (15w). (BD) Whole orgn mounts of BAs (B) isodca (C) n CA7 sulfte (D) in cecum. n=57 mice/group; D, onor smple; MCA, muricholic ci; CA, cholic ci; HDCA, hyoeoxycholic ci; DCA, eoxycholic ci; T, turineconjugte species.
7 cteri Cynob 4C_2 CONVM (2 weeks) CONVH (2 weeks) t r g b2 b1 b Verrucomic obcteri robi Actin Te iiverrucomicrob n e ric obcter i ute Cori M o e s llic ute 7 c 9 s Ep 8 b sil f b6 b5 b4 b3 e b7 h c es B cte ro B ie q cte p t ro o n s ii m l k i j 1 illi Bc w z v y u x teri Alphproteobc bc teri teo bcteri E r proteo ysip Pro elo Bet c7 2 tric ri cte c6 hi 1 c9 b eo c5 ot 6 c8 c4 pr c 3 3 c2 on 4 5 c b9 1 b8 : Alercreutzi b: Collinsell_erofciens c: Coriobctericee : Coriobcteriles e: Bcteroies f: Bcteroies_ovtus g: Brnesiellcee h: Butyricimons i: Ooribcter j: Ooribctercee k: Prbcteroies l: Prbcteroies_istsonis m: Porphyromoncee n: Prevotell o: Prevotell_copri p: Prevotellcee q: Rikenellcee r: S24_7 s: Bcteroiles t: YS2 u: Stphylococcus v: Stphylococccee w: Bcillles x: Lctobcillus y: Lctobcillcee z: Lctobcillles : Christensenellcee 1: Clostriicee 2: Dehlobcterium 3: Dehlobctericee 4: Anerofustis 5: Eubctericee 6: Bluti 7: Bluti_obeum 8: Coprococcus 9: Dore b: Roseburi b1: Ruminococcus b2: Lchnospirchee b3: Mogibctericee b4: rc4_4 b5: Feclibcterium_prusnitzii b6: Ruminocuccus b7: Ruminocucccee b8: Phscolrctobcterium b9: Vellionellcee c: Clostriiles c1: Allobculum c2: Coprobcillus c3: Eubcterium _olichum c4: cc115 c5: Erysipelotrichcee c6: Erysipelotrichles c7: RF32 c8: Suturell c9: Alcligencee : Oxlobcter_formigenes 1: Oxlobctercee 2: Burkholeriles 3: Bilophil 4: Desulfovibrio 5: Helicobctercee 6: Cmpylobcterles 7: RF39 8: Akkermnsi_muciniphil 9: Verrucomicrobicee e: Verrucomicrobiles Supplementl Figure S2. Clogrm representing OTUs of sttisticl n biologicl ifference between cecum microbiot of CONVM n CONVH mice fter 2 weeks of coloniztion. Ech circle imeter is proportionl to OTU s bunnce. n=6 smples/group.
8 v u t s q r Bc tero ie Bc tes ter oi i x p n o m l j c3 b3 b8b7 b6b5 b4 9 b b2 b1 c4 2 c1c b c 9 c6 8 7 c5 7 c CONVM (15 weeks) CONVH (15 weeks) teri nobc Cy _2 4C y i cill w 1 B 6 3 z k i g f Verrucomicrob i T e n Verrucomi eric c r o b i e ute cterie Mol s inob l Act i c u tes G f3 m m f2 p r h e c b f6 f5 f4 2 f f1 e8 e9 teri teobc oteobcter r p h p l A i pro Bectteri 8 3 e ri eob cte rot e 7 b ltp e7 e eo e 4 e1 ot D 1 e6 3 e4 e5 c8 c9 : Bifiobcterium b: Bifiobcterium_longum c: Bifiobctericee : Bifiobcteriles e: Alercreutzi f: Collinsell_erofciens g: Coriobctericee h: Bcteroies i: Bcteroies_ovtus j: Bcteroies_uniformis k: Bcteroicee l: Brnesiellcee m: Butyricimons n: Ooribcter o: Ooribctercee p: Prbcteroies q: Prbcteroies_istsonis r: Porphyromoncee s: Prevotell t: Prevotell_copri u: Prevotellcee v: Rikenellcee w: S24_7 x: Bcteroiles y: YS2 z: Stphylococcus : Stphylococccee 1: Bcillles 2: Enterococcus 3: Enterococccee 4: Lctobcillus 5: Lctobcillcee 6: Lctobcillles 7: Christensenellcee 8: Cnitus_rthromitus 9: Clostriicee b: Dehlobcterium b1: Dehlobctericee b2: Anerofustis b3: Eubctericee b4: Bluti b5: Bluti_obeum b6: Coprococcus b7: Coprococcus_eutctus b8: Dore b9: Dore_formicigenerns c: Ruminococcus c1: Ruminococcus_gnvus c1: Ruminococcus_torques c3: Lchnospirchee c4: Mogibctericee c5: rc4_4 c6: Peptococccee c7: Feclibcterium_prusnitzii c8: Ruminocuccus c9: Ruminocucccee : Phscolrctobcterium 1: Vellionellcee 2: Clostriiles 3: Allobculum 4: Coprobcillus 5: Eubcterium _olichum 6: cc115 7: Erysipelotrichcee 8: RF32 9: Suturell e: Alcligencee e1: Oxlobcter_formigenes e2: Oxlobctercee e3: Burkholeriles e4: Bilophil e5: Desulfovibrio e6: Desulfovibrioncee e7: Desulfovibrionles e8: Enterobctericee e9: Enterobcteriles f: Aneroplsm f1: Aneroplsmtcee f2: Aneroplsmtles f3: RF39 f4: Akkermnsi_muciniphil f5: Verrucomicrobicee f6: Verrucomicrobiles Supplementl Figure S3. Clogrm representing OTUs of sttisticl n biologicl ifference between cecum microbiot of CONVM n CONVH mice fter 15 weeks of coloniztion. Ech circle imeter is proportionl to OTU s bunnce. n=61 smples/group.
9 A B C Primry BAs Seconry BAs CONVM 2 w CONVH 2 w CONVH D2 2 w D E F CONVM 15 w CONVH 15 w CONVH D2 15 w Supplementl Figure S4. Reltive mounts of primry n seconry BAs in cecum of mice colonize with mouse or humn microbiot. (AF) Primry n seconry BAs in cecum fter 2 weeks (AC) or 15 weeks (DF) coloniztion with mouse microbiot (A n D), humn microbiot from the first humn onor (CONVH) (B n E) or the secon humn onor (CONVH D2) (C n F). Primry bile cis, TCA, CA, TCDCA, CDCA, TαMCA, αmca, TβMCA, βmca, TUDCA, UDCA); seconry BAs, DCA, isodca, HCA, HDCA, LCA, MDCA, ωmca,
10 A B C Conjugte BAs Unconjugte BAs CONVM 2 w CONVH 2 w CONVH D2 2 w D E F CONVM 15 w CONVH 15 w CONVH D2 15 w Supplementl Figure S5. Reltive mounts of conjugte n unconjugte BAs in cecum of mice colonize with mouse or humn microbiot. (AF) Conjugte n unconjugte BAs in cecum fter 2 weeks (AC) or 15 weeks (DF) coloniztion with mouse microbiot (CONVM) (A n D), humn microbiot from the first humn onor (CONVH) (B n E) or the secon humn onor (CONVH D2) (C n F). Conjugte BAs, TCA, TCDCA, TDCA, THDCA, TαMCA, TβMCA, TωMCA, TUDCA; unconjugte BAs, CA, CDCA, DCA, isodca, HCA, HDCA, LCA, αmca, βmca, ωmca, MDCA, UDCA.
11 A B Concentrtion of BAs (mm) TCA TDCA GCA CA TCA h TCA 24 h DCA Concentrtion of BAs (mm) TCA TDCA GCA CA DCA CA h CA 24 h Supplementl Figure S6. Metbolism of TCA n CA by humn microbiot in vitro. (A) Concentrtion of BAs in suspensions of humn fecl smples incubte in BHI meium with TCA (A) or CA (B) for 24 hours. Men vlues ± SEM re plotte; n=2 smples for ech time point.
12 A FXR gonist/ntgonist rtio in liver 2 w 15 w B FXR gonist/ntgonist rtio in gllbler w 15 w b GF CONVM CONVH C FXR gonist/ntgonist rtio in portl vein w 15 w D FXR gonist/ntgonist rtio in ven cv w 15 w Supplementl Figure S7. Chnges in the rtio between FXR gonists n ntgonists fter coloniztion. (AD) Rtio between FXR gonists n ntgonists in liver (A), gllbler (B), portl vein (C) n cvl vein (D). Men vlues ± SEM re plotte. n= 49 smples/group; P<.5, b P<.1, c P<.1, P<.1 inicte ifferences vs GF with ANOVA n Dunnett s multiple comprisons test. FXR gonists, TCA, TCDCA, TDCA, TLCA, CA, CDCA, DCA, LCA; FXR ntgonists, TαMCA, TβMCA.
13 Supplementl Tble S1. Bile ci (BA) content in liver. Amounts of BAs in the whole orgn for ech group of mice. BAs >.1% bunnce re inclue. BAs (nmol/orgn), reltive liver weight (mg/g boy weight) n boy weight (g) re presente s men ± SEM; n = 59 smples/group; P<.5, b P <.1, c P <.1, P <.1 inicte ifferences vs GF mice with ANOVA n Tukey s multiple comprisons test. Liver BAs GF CONVM (2 w) CONVH ( 2w) CONVM (15 w) CONVH (15w) TMCA.19 ±.3.21 ±.4.19 ±.4.8 ±.2.7 ±.1 TbMCA.77 ± ± ± ±.21 b.279 ±.44 TwMCA.77 ± ±.7.1 ±. TCA.291 ± ± ± ± ±.32 TUDCA.4 ±.1.3 ±.1.5 ±.1.2 ±..2 ±. THDCA.3 ±.1 trces TCDCA.8 ±.1.5 ±.1.5 ±.1.2 ±. b.2 ±. c TDCA.9 ±.2.2 ±.1.4 ±.1.1 ±. bmca.3 ±.1 trces.4 ±.2 trces.3 ±.1 CA trces Totl BA 1.95 ± ± ± ±.56 b.463 ±.77 b Reltive liver weight 44. ± ± ± ± ±.71 Boy weight 29.2 ± ± ± ± 2.1 c 29.3 ±.8
14 Supplementl Tble S2. Bile ci (BA) content in gllbler. Amounts of BAs in the whole orgn for ech group of mice. BAs >.1% bunnce re inclue. BAs (nmol/orgn) n reltive weights (mg/g boy weight) re presente s men ± SEM; n = 59 smples/group; P<.5, b P <.1, c P <.1, P <.1 inicte ifferences vs GF mice with ANOVA n Tukey s multiple comprisons test. Gllbler BAs GF CONVM (2 w) CONVH (2 w) CONVM (15 w) CONVH (15 w) TMCA ± ± ± 9.34 b ± ± 9.9 TbMCA ± ± b ± b ± ± TwMCA ± c ± ± 3.56 TCA ± ± ± b ± ± TUDCA ± ± ± 3.53 c ± ± 1.1 THDCA ± ± 5.77 b ± 1.26 TCDCA ± ± 2.48 b ± 1.4 b ± 6.6 trces TDCA 4.5 ± ± 9.44 trces bmca 24.3 ± trces wmca 2.98 ± 1.55 b CA trces ± b Totl BA ± ± c ± 9.31 c ± ± b Reltive gllbler weight 2.85 ± ±.8.67 ± ± ±.15
15 Supplementl Tble S3. Bile ci (BA) content in cecum. Amounts of BAs in the whole orgn for ech group of mice. BAs >.1% bunnce re inclue. BAs (nmol/orgn) n reltive weights (mg/g boy weight) re presente s men ± SEM; n = 59 smples/group; P<.5, b P <.1, c P <.1 P <.1 inicte ifferences vs GF mice with ANOVA n Tukey s multiple comprisons test. Cecum BAs GF CONVM (2 w) CONVH (2 w) CONVM (15 w) CONVH (15 w) TMCA ± ± ± ± ± 5.5 TbMCA ± ± c ± c ± 31. c ± c TwMCA 27.2 ± ± 7.57 trces TCA ± ± ± 6.27 b ± b ± 16.5 TUDCA 3.13 ±.35 trces.68 ±.9 c trces THDCA 1.65 ±.73 trces trces TCDCA 2.33 ±.37 trces TDCA 3.5 ± 1.39 trces 1.67 ±.3 MCA 3.5 ± ± ± ±.47 bmca 23.1 ± ± ± ± b wmca ± ± ± ±.89 CA ± ± ± b ± HCA trces UDCA.98 ± ± ±.38 c 1.33 ±.22 MDCA trces trces trces HDCA 13.2 ± 3.52 c 2.58 ± ± 1.91 c 5. ±.74 CDCA.88 ± 1.19 trces trces DCA ± c 22.9 ± ± ± 11.1 LCA 5.63 ±.92 c 4.24 ± ±.82 b 1.81 ±.53 Totl BA ± ± ± ± ± Reltive cecum weight ± ± ± ± ± 1,8
16 Supplementl Tble S4. Bile ci (BA) content in portl vein. BA concentrtions (nmol/ml) in portl vein serum for ech group of mice. BAs >.1% bunnce re inclue n presente s men ± SEM; n = 59 smples/group; P<.5, b P <.1, c P <.1, P <.1 inicte ifferences vs GF mice with ANOVA n Tukey s multiple comprisons test. Portl vein BAs GF CONVM (2 w) CONVH (2 w) CONVM (15 w) CONVH (15 w) TMCA 6.95 ± ± ± ± ±.44 TbMCA ± ± c ± 15. b 38.3 ± c ± c TwMCA 1.4 ± 3.43 b 1.68 ± 3.28 b trces TCA ± ± ± ± ± 7.66 TUDCA 2.1 ± ±.11 b.69 ± ±.1 b.56 ±.1 c THDCA.32 ±.8 trces.46 ± ±.6 TCDCA 1.47 ± ±.5 b.53 ±.9 trces.34 ±.6 c TDCA.76 ±.18 trces.85 ±.2 b.24 ±.4 MCA.24 ±.5.52 ±.16 trces bmca 1.8 ± ± ± ± 1.82 wmca 1.37 ± ±.86 CA 7.6 ± ± ± 5.65 b 6.57 ± 1.43 UDCA trces.55 ±.1.26 ±.5 HDCA.13 ±.4.27 ±.3 CDCA trces DCA 1.27 ± ± ± ±.5 Totl BA ± ± b ± ± b ± b
17 Supplementl Tble S5. Bile ci (BA) content in cvl vein. BA concentrtions (nmol/ml) in cvl vein serum for ech group of mice. BAs >.1% bunnce re inclue n presente s men ± SEM; n = 59 smples/group; P<.5, b P <.1, c P <.1, P <.1 inicte ifferences vs GF mice with ANOVA n Tukey s multiple comprisons test. Cvl vein BAs GF CONVM (2 w) CONVH (2 w) CONVM (15 w) CONVH (15 w) TMCA.37 ±.13.2 ±.1.9 ±.4.4 ±.1.5 ±.2 TbMCA ± ± ±.67.3 ± ±.49 TwMCA.8 ±.2.14 ±.4 TCA 7.5 ± ± ± ± ±.52 TUDCA.6 ±.2 trces.1 ±. trces.1 ±. THDCA trces.1 ±..1 ±..1 ±..1 ±. TCDCA.7 ±.2.1 ±..2 ±..1 ±. TDCA.3 ±.1.1 ±.1.3 ±.1.1 ±. MCA.2 ±.1.2 ±.1.1 ±. bmca.9 ±.3.8 ±.2.33 ± ±.22.8 ±.23 wmca.11 ±.4.44 ±.12.1 ±. CA.19 ±.5.12 ±.8.99 ±.27 b.28 ±.8 UDCA trces.1 ±.1.3 ±.1.2 ±.1 MDCA trces HDCA.1 ±. trces.1 ±..1 ±. CDCA trces trces DCA.7 ±.2.2 ±.1.13 ±.1.2 ±. Totl BA 22.2 ± ± ± ± ± 1.25
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