Fig 2: Antibiotic treatment improves metabolism in adipose tissue Napepld-deleted mice.(a) Total body-weight gain (g; n=5). (b) Adiposity index (% Total body weight) after long-term antibiotic treatment (n=5). (c) Plasma glucose (mg dl−1) profile and (d) the mean area under the curve (AUC) measured between 0 and 120 min after glucose loading (mg l−1 min−1; n=5). (e) Plasma insulin levels at 30 min before and 15 min after glucose loading (μg l−1) (n=5). (f) Insulin resistance index determined by multiplying the AUC of blood glucose by the AUC of insulin. Data are presented as mean±s.e.m. Data with ‘*’ are significantly different (P<0.05) according to the unpaired two-tailed Student t-test, or following a two-way statistical analysis in c. Data with ‘**’ indicates a significant difference (P<0.01) versus time point −30 min in cKO mice in e.

Fig 3: Effect of maternal and/or offspring exposure to a free-choice palatable (P) diet on the relative protein levels of: (A), GRIA1 (B), mGLUR5 (C), CB1 (D), CB2 (E), DAGLα (F), DAGLβ (G), MAGL (H), NAPE-PLD (I) and FAAH (J) in the prefrontal cortex of male and female offspring in adulthood. Representative immunoblots (K). Data are expressed as the mean ± S.E.M. (n = 4). Student’s t test: */**/*** p < 0.05/0.01/0.001 vs. CC males or CC females; # p < 0.05 vs. CP males or CP females. MD: maternal diet effect; PD: postnatal diet effect; MDxPD: maternal and postnatal diets interaction; SxMDxPD: sex, maternal diet and postnatal diet interaction.

Fig 4: Adipose tissue Napepld deletion alters adipose and whole-body lipid metabolism.(a) Plasma triglyceride content (mg dl−1; n=20–27). (b) Plasma cholesterol content (mg dl−1; n=20–27). (c) Plasma non-esterified fatty acid (NEFA) levels (mM; n=20–27). These data (a–c) correspond to the results of three independent experiments. (d) SAT prostaglandin content measured by LC-MS/MS in fmol per mg tissue (n=10). (e) SAT eicosanoid content measured by LC-MS/MS in fmol per mg tissue (n=10). (f) SAT ceramide content measured by ESI-MS/MS in pmol per mg tissue (n=10). See also Supplementary Table 1. Data are presented as mean±s.e.m. Data with different superscript letters or symbols are significantly different (P<0.05) according to post-hoc one-way analysis of variance. ‘*’ indicates a significant difference (P<0.05) versus WT-CT, and ‘#’ indicates a significant difference (P<0.05) versus WT-CT and cKO-CT according to post-hoc one-way analysis of variance.

Fig 5: Specific deletion of Napepld in adipose tissue.(a) Representative adipose tissue immunoblot of NAPE-PLD and β-actin in WT mice and cKO mice. (b) mRNA relative expression of Napepld in different adipose tissue deposits (subcutaneous, visceral, epididymal and brown adipose tissue—SAT, VAT, EAT and BAT), as well as in liver, colon and tibialis muscle under CT diet and HFD in cKO mice and WT mice (n=20–27). These data (a,b) correspond to the results of three independent experiments. Data with ‘*’ indicate a significant difference (P<0.05) versus WT-CT or WT-HFD according to the unpaired two-tailed Student’s t-test. (c) SAT levels of AEA, PEA, OEA and SEA (pmol per g tissue) measured by HPLC-MS (n=6–10). Data are presented as the mean±s.e.m. Data with different superscript letters are significantly different (P<0.05) according to post-hoc one-way analysis of variance statistical analysis.