ARTICLES ●cod oTU 32 Palndibacter oTU 897 oTU 3 orphyromonadaceae aludibacter oTU 998 Osciibacter OTU 15 Deferribacteraceae oTu 7 Porphyromonadaceae Parabacteroides OT Ds1(52%) Clostridium xIva OTU 68 小导4杂P98 Genera Mean relative abundance (% Mean relative abundance(%) t Clostridiales ■Unc. Porphyromonas■Uc Anaerotruncus Uncl. Lachnospiraceae UncL. ■Unc. Erysipelotrichac.■uncL zEz9 Cold g8E8型 Food intake Feces Fracto k Figure 1 BAT activation alters the gut microbiome, lipoprotein levels and cholesterol uptake (a-e)MDS plot of weighted UniFrac distance(a), mean relative sundance of gut microbiota on family and genus level( b), hierarchical clustering of significant altered oTUs (c), as well as alpha diversity presented at the (w), n=9 mice)or 6C(cold (c), n=5 mice). (f)Food intake and amount of feces per 24 h period (n= 6 mice per group). (g) Plasma triglyceride (g)and cholesterol(Chol)levels(n= 5 mice per group).(h, i) Profiles of triglyceride-rich lipoproteins(TRLs)and cholesterol-rich LDL and hdl by measurement of triglycerides(h)and cholesterol (i)in FPLC fractions, respectively (pooled plasma samples of n=5 mice per group).( k)Organ uptake of 3H-radiolabeled P<0.05, ""P<0.01.*P<0.001: n.s., not significant; by pairwise Wilcoxon rank-sum test(d, e)or unpaired two-tailed Student's -test (f,gk-mgWAT holesterol 4 h after oral gavage, expressed as fold induction (i or as percentage of injected dose for liver and small intestine ( k)(n=7 mice per group). ing guinal WAT; epiwAT, epididymal WAT; ScBAT, subscapular BAT; iBAT, interscapular BAT: deBAT, deep cervical BAT. Throughout, data are mean +se© 2017 Nature America, Inc., part of Springer Nature. All rights reserved. a r t i c l e s nature medicine advance online publication  MDS2 (20%) MDS1 (52%) Warm Cold Warm Warm *** *** *** *** ** *** *** *** * * Food intake Feces Cold Cold Warm Cold Warm Cold Warm Cold TRL Warm HDL LDL Cold Warm Cold Warm Cold Warm Cold Warm Cold Warm Warm Cold Warm Families Mean relative abundance (%) Porphyromonadaceae Mean relative abundance (%) Parabacteroides Uncl. Porphyromonad. Uncl. Ruminococc. Mucispirillum Anaerotruncus Other Uncl. Marinilabiliac. Uncl. Clostridiales Uncl. Synthrophom. Uncl. Lachnospiraceae Uncl. Erysipelotrichac. Genera Cold Cold log10 scaled abundance Family Lachnospiraceae Lachnospiraceae Porphyromonadaceae Porphyromonadaceae Porphyromonadaceae Ruminococcaceae Ruminococcaceae Erysipelotrichaceae Erysipelotrichaceae Other 30 2.0 160 3 2 1 0 120 80 40 0 1.5 1.0 0.5 0.0 20 10 0 5 200 3 10 8 6 4 2 0 2 1 0 0 10 20 Fraction 30 0 10 20 Fraction 30 150 100 TG Chol 50 0 4 3 2 1 0 5 4 3 2 1 0 n.s. n.s. ** ** Observed genera Amount (g/24 h) Organ uptake (cpm/organ, fold) Organ uptake (% of given dose) Concentration (mg/dl) TG (mg/dl) Chol (mg/dl) Shannon (genera) Observed OTUs Shannon (OTU) Marinilabiliaceae Clostridiales Synthrophomonadaceae Deferribacteraceae Deferribacteraceae Porphyromonadaceae Lachnospiraceae w6 Plasma Liver Spleen Kidney Muscle epiWAT ingWAT scBAT iBAT dcBAT Liver Duodenum Jejunum Ileum Intestine total w1w2 w9 w3 w7 w5w4 w8c2 c3 c1 c4c5 Genus OTU OTU_32 OTU_897 OTU_3 OTU_998 OTU_15 OTU_23 OTU_7 OTU_1 OTU_68 incertae sedis Paludibacter Paludibacter Paludibacter Oscillibacter Allobaculum Mucispirillum Parabacteroides Clostridium XIVa –2 –1 0 1 2 0.10 0.1 0.2 –0.10 –0.2 –0.1 0.05 –0.05 0 0 a b d f j k g h i c Figure 1 BAT activation alters the gut microbiome, lipoprotein levels and cholesterol uptake. (a–e) MDS plot of weighted UniFrac distance (a), mean relative abundance of gut microbiota on family and genus level (b), hierarchical clustering of significant altered OTUs (c), as well as alpha diversity presented at the genus level (d) or at the OTU level (e), on the basis of 16S-rRNA-encoding sequences in feces collected from mice that were housed at thermoneutrality (warm (w), n = 9 mice) or 6 °C (cold (c), n = 5 mice). (f) Food intake and amount of feces per 24 h period (n = 6 mice per group). (g) Plasma triglyceride (TG) and cholesterol (Chol) levels (n = 5 mice per group). (h,i) Profiles of triglyceride-rich lipoproteins (TRLs) and cholesterol-rich LDL and HDL by measurement of triglycerides (h) and cholesterol (i) in FPLC fractions, respectively (pooled plasma samples of n = 5 mice per group). (j,k) Organ uptake of 3H-radiolabeled cholesterol 4 h after oral gavage, expressed as fold induction (j) or as percentage of injected dose for liver and small intestine (k) (n = 7 mice per group). ingWAT, inguinal WAT; epiWAT, epididymal WAT; scBAT, subscapular BAT; iBAT, interscapular BAT; dcBAT, deep cervical BAT. Throughout, data are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001; n.s., not significant; by pairwise Wilcoxon rank-sum test (d,e) or unpaired two-tailed Student’s t-test (f,g,j,k)