ARTICLES b d Wam● Cold o Warm EZ。 Cold ez ● Coldez 意16n邑 6i是 TG Chol CBA ng) ug owam。WamE g owam。 Warm EZ o Warm o Warm EZ ● Cold eZ ●cod● Cold EZ ●cod● Cold ez 芦芹 g28·88:-201·是g8目e量8e.。201 i..dit △Wam △wam ◆ Warm EZ口coEz ◆ Warm Chol口 Cold cho ◇cp7 bl warm口 ◇ 8△◇ ◇ 0.00● ◇ 0051◇ °公°△ 0.10 △ -0.10-0050000050.10 .3-02.10.00.102 MDs1(37%) Ds1(52%) s1(80%) △AAV-GFP●AAv-Cyp7b △ AAV-GFP● AAV-Cyp7b1 △Mdr2wam●Md2'co 0.1 ● -0030.000.03006 10-0050.000.05 .1000.1 MDs1(43%) Figure 5 Cold-induced bile acid excretion determines the composition of the gut microbiome (a-h) Plasma lipids (a), lipoprotein profiles of poole lasma(b), hepatic lipid content (c), hepatic expression of HMG-CoA reductase( Hmgcr) and Ldlr mRNAs(d), hepatic UBA and CBA levels (e), total fecal UBA and CBA levels(f), as well as fecal unconjugated (g) and conjugated and unconjugated a-MCA (h) bile acid species, as determined in arm-housed and cold-housed mice that were treated without or with ezetimibe(Ez)(for a-d, f -h, n=8 mice per group; for e, warm-housed: no ez warm), n=6 mice: +EZ, n=7 mice; cold-housed: no EZ(cold), n=8 mice: +EZ, n=8 mice). (i-n)MDS plots of weighted UniFrac-distance-based 6s rRNA-encoding sequence analysis of fecal samples from warm-housed and cold- housed mice that were treated without or with Ez (warm-housed: no EZ (warm), n=8 mice; +EZ, n= 10 mice; cold-housed: no EZ (cold), n=7 mice; +EZ, n=9 mice)(i), warm-housed and cold- housed mice that were fed a chow diet supplemented without or with cholesterol (warm-housed, no added Chol (warm)n=8 mice; +Chol, n=8 mice; cold- housed: no added 4 mice per group)(k), mice that were infected with either AAV-GFP or AAV-Cyp7bl and housed at 30C (AAV-GFP, n= 7 mice; AAv-Cyp7bl, n=5 mice)(I)or 22C(AAV-GFP, n=7 mice per group)(m), and warm-housed and cold- housed Mdr2--mice(warm-housed, n=7 mice; cold-housed n=8 mice)(n). Throughout, data are mean ts.e. m. 'P<0.05, *P<0.01,"*P<0.001; by two-way ANovA (a-h)© 2017 Nature America, Inc., part of Springer Nature. All rights reserved. Ar t i c l e s  advance online publication nature medicine Warm Cold Warm EZ Cold EZ 8 6 4 2 0 Gene expression (fold) Bile acids (ng/mg, fold) Bile acids (µg/mg) Bile acids (ng/mg) Bile acids (ng/mg, fold) MDS2 (19%) MDS2 (19%) MDS2 (19%) MDS2 (21%) MDS2 (17.1%) MDS2 (24%) Liver lipids (per mg/protein) Cholesterol (mg/dl) Lipids (mg/dl) 2.0 *** *** ** *** ** *** * 1.5 1.0 0.5 0.0 Fraction TG (mg) Chol (µg) 40 30 20 10 0 0 10 20 30 250 200 150 100 50 0 TG UBA UBA CBA CBA CA β-MCA CDCA/DCA UDCA ω-MCA α-MCA T-α/β-MCA THDCA TUDCA TDCA TCDCA TLCA TCA Chol 20 15 10 5 0 5 4 3 2 1 0 10 8 6 4 2 0 100 80 60 40 20 0 200 150 250 100 50 0 0.10 0.05 0.00 –0.05 0.10 0.05 0.00 –0.05 –0.10 0.2 0.1 0.0 –0.1 0.05 0.00 –0.05 0.10 0.05 0.00 –0.05 –0.10 –0.3 –0.2 –0.1 0.0 0.1 0.2 –0.2 –0.1 0.0 0.1 0.2 –0.1 0.0 0.1 0.2 –0.10 –0.05 0.00 0.05 0.10 –0.03 0.00 0.03 0.06 MDS1 (37%) –0.10 –0.05 0.00 0.05 MDS1 (30%) MDS1 (43%) MDS1 (52%) MDS1 (80%) MDS1 (42%) Hmgcr Ldlr Warm Warm EZ Cold Cold EZ Warm Cold Warm EZ Cold EZ Warm Warm EZ Cold Cold EZ Warm Warm EZ Cold Cold EZ Warm Warm EZ Cold Cold EZ Warm Cold AAV–GFP AAV–Cyp7b1 AAV–GFP AAV–Cyp7b1 Mdr2–/– warm Mdr2–/– cold Warm EZ Cold EZ Warm Cold Warm Chol 30 °C 22 °C Cold Chol WT warm WT cold Cyp7b1–/– warm Cyp7b1–/– cold 0.050 –0.050 0.025 –0.025 0.000 a b c d e f g h i j k l m n *** *** *** *** * *** *** *** *** * *** *** * *** ** *** *** *** *** *** * *** ***** *** * ***** *** **** * * ** ** * ** **** *** *** *** ** ** ** ** ** ** ** ** ** *** ** *** *** *** *** Figure 5 Cold-induced bile acid excretion determines the composition of the gut microbiome. (a–h) Plasma lipids (a), lipoprotein profiles of pooled plasma (b), hepatic lipid content (c), hepatic expression of HMG-CoA reductase (Hmgcr) and Ldlr mRNAs (d), hepatic UBA and CBA levels (e), total fecal UBA and CBA levels (f), as well as fecal unconjugated (g) and conjugated and unconjugated α-MCA (h) bile acid species, as determined in warm-housed and cold-housed mice that were treated without or with ezetimibe (EZ) (for a–d,f–h, n = 8 mice per group; for e, warm-housed: no EZ (warm), n = 6 mice; +EZ, n = 7 mice; cold-housed: no EZ (cold), n = 8 mice; +EZ, n = 8 mice). (i–n) MDS plots of weighted UniFrac-distance-based 16S rRNA-encoding sequence analysis of fecal samples from warm-housed and cold-housed mice that were treated without or with EZ (warm-housed: no EZ (warm), n = 8 mice; +EZ, n = 10 mice; cold-housed: no EZ (cold), n = 7 mice; +EZ, n = 9 mice) (i), warm-housed and cold-housed mice that were fed a chow diet supplemented without or with cholesterol (warm-housed, no added Chol (warm) n = 8 mice; +Chol, n = 8 mice; cold-housed: no added Chol (cold), n = 7 mice; +Chol, n = 8 mice) (j), warm-housed and cold-housed WT control and Cyp7b1−/− mice (WT:, n = 5 mice per group; Cyp7b1−/−, n = 4 mice per group) (k), mice that were infected with either AAV-GFP or AAV-Cyp7b1 and housed at 30 °C (AAV-GFP, n = 7 mice; AAV-Cyp7b1, n = 5 mice) (l) or 22 °C (AAV-GFP, n = 7 mice per group) (m), and warm-housed and cold-housed Mdr2−/− mice (warm-housed, n = 7 mice; cold-housed, n = 8 mice) (n). Throughout, data are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001; by two-way ANOVA (a–h)