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. 2022 Jun 24;10:96. doi: 10.1186/s40168-022-01264-5

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Microbiota depletion impairs weight loss and metabolic improvements resulting from RYGB surgery in diet-induced obesity. a–f Relative body weight change (in %; average body weight (øBW) per group: RYGB(ABx) week (W)0: 385.6 g, W5: 454.3 g; DIO W0: 681.3 g, W5: 762.3 g; RYGB W0: 415.3 g, W5: 437.7 g; # DIO vs. RYGB; * RYGB vs. RYGB(ABx)) (a), final body composition with lean and fat mass (in g), b cumulative energy intake (in kcal) (c), cumulative energy intake of high fat diet (HFD) and standard chow (SC) (in g) (d), energy assimilation (in kJ/24 h) (e), oral glucose-tolerance tests (oGTT) (f), and insulin-sensitivity assessed by insulin-tolerance tests (ITT) (g) in RYGB-operated rats with 35-day antibiotic-treatment (ABx) compared to their respective controls (RYGB-operated rats without ABx). h–k Fasting plasma triglyceride concentrations (in ng/μl) (h), representative images (at least 4 images per group) of liver Oil Red O staining (scale bars 200 μm) with quantification of hepatic lipid accumulation (in %) (i), and relative mRNA expression of hepatic pro- and anti-inflammatory cytokines (j) in RYGB(ABx) compared to RYGB rats. Plasma cytokine concentrations in RYGB(ABx) and RYGB rats (in pg/ml) (k). l-n Representative images of H&E and UCP1 staining in BAT (scale bars 200 μm) (l), total energy expenditure (TEE; in kcal/day) (m) and infrared images of representative RYGB(ABx) or RYGB rats after 6 h of cold-exposure (n). Mouse primary brown adipocytes were treated for 6 h with 50% (vol/vol) of rat serum from RYGB(ABx) or RYGB rats (or with PBS as control). Oxygen consumption rate was measured on a respirometry Seahorse XF Cell Mito Stress Test (Agilent) as described in the “Methods” section (o). Experiments were performed in three independent cohorts. Data are mean ± s.e.m; n = 3–8 animals per group with pooled data from 2 to 3 independent experiments. *,#P < 0.05, **,## P < 0.01, ***,### P <0.001 as assessed by unpaired Student’s t test (for two groups) or two-way ANOVA (multiple groups) with Tukey correction for multiple testing