Journal: Molecular Metabolism

Article Title: Differential cell type-specific function of the aryl hydrocarbon receptor and its repressor in diet-induced obesity and fibrosis

doi: 10.1016/j.molmet.2024.101963

Figure Lengend Snippet: AhRR-deficient mice are partially resistant to diet-induced obesity. ( A ) Body weight of wild-type (WT) and AhRR-deficient mice fed CD or HFD for 14 weeks; n = 8 (CD WT), n = 6 (HFD WT), n = 5 (CD, HFD AhRR −/− ). ( B ) Blood glucose levels during a glucose tolerance test -GTT- (left panel) and insulin tolerance test -ITT- (right panel) after 12 weeks on HFD; n = 11 (GTT WT), n = 10 (GTT AhRR −/− ), n = 5 (ITT WT, ITT AhRR −/− ). ( C ) Total serum cholesterol levels after 14 weeks on HFD; n = 9 (WT), n = 7 (AhRR −/− ). ( D ) Analysis of covariance (ANCOVA) (non-linear fit) of oxygen consumption/body weight (BW); n = 8 for both groups. ( E ) Total body fat mass assessed by NMR; n = 8 for both groups. ( F ) WATg weight; n = 8 for both groups. ( G ) Representative H&E staining. Scale bars, 80 μm (left panel). Right, quantification of adipocyte size of the WATg of HFD mice; n = 5 (WT), n = 4 (AhRR −/− ). ( H ) Liver weight; n = 8 (CD WT, CD AhRR −/− , HFD AhRR −/− ), n = 7 (HFD WT). ( I ) Representative H&E (left panel) and Sirius Red (right panel) staining of liver sections. Scale bars, 100 μm. ( J ) Representative Oil Red O (ORO) and Hemalaun staining of liver sections of WT and AhRR-deficient mice after HFD. Scale bars, 80 μm (left panel). Right, quantification of ORO stain, n = 3 (CD WT), n = 8 (HFD WT), n = 6 (HFD AhRR −/− ). ( K ) Serum Alanine transaminase (ALT) concentration of WT and AhRR-deficient mice fed either CD or HFD for 14 weeks; n = 4 for all groups. ( L ) Body weight of WT and AhRR-deficient mice fed a HFD supplemented with or without indole-3 carbinole (I3C) for 14 weeks; n = 20 (WT HFD), n = 15 (WT HFD + I3C), n = 15 (AhRR −/− HFD), and n = 14 (AhRR −/− HFD + I3C). ( M ) Representative ORO and Hemalaun staining of liver sections of WT and AhRR −/− mice after HFD ± I3C. Scale bars, 100 μm (left panel). Right, ORO staining quantification of liver sections; n = 10 (WT CD, HFD), n = 7 (WT HFD + I3C), n = 5 (AhRR −/− HFD), n = 9 (AhRR −/− HFD + I3C). ( N ) z-scored intensity values of representative TCA-metabolites found in plasma of mice after 14 weeks of CD or HFD ± I3C; n = 3–5. ∗p < 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001. Significance was determined using unpaired two-tailed t-tests (A, B, C, G) and one-way analysis of variance (ANOVA) with Tukey's multiple-comparison test (E, F, H, K and M) or with Dunnett's multiple comparisons test (J). Two-way analysis of variance (ANOVA) with Tukey's multiple-comparison test (L). Data are mean ± s.e.m.

Article Snippet: For dietary intervention studies, mice were exposed to a purified diet or with 2 g/kg indole-3-carbinole (I3C)-supplemented control diet (CD/CD+I3C, 13 kJ% Fat, Cat No.: D12450 LS ssniff EF) or a purified or with 2 g/kg I3C-supplemented high fat diet (HFD/HFD+I3C, 60 kJ% fat, Cat. No.: D12492 ssniff EF) ad libitum .

Techniques: Staining, Concentration Assay, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: PI(18:1/18:1) is a SCD1-derived lipokine that limits stress signaling

doi: 10.1038/s41467-022-30374-9

Figure Lengend Snippet: NIH-3T3 fibroblasts were treated with vehicle, TNFα (10 ng/ml), STS (0.3 µM), CHX (20 µg/ml), ETO (10 µM), TPG (2 µM), VAL (10 µM), MC (10 µM), or I3M (10 µM) or were serum starved (Serum) for the indicated period of time. a , d – f Cellular proportion of MUFAs in PI and PI(18:1/18:1) (left to right (LTR) P = 0.0000004, 0.00000007, 0.0000007) ( a ), PC (LTR P = 0.9999999986, 0.000002, 0.0000003) ( d ), PE (LTR P = 0.99991, 0.000000007, 0.99999992) ( e ), and PS (LTR P = 0.99992, 0.99998, 0.99997) ( f ); MUFAs: 16:1, 18:1. Data of ( a ), ( d ) is identical to w/o in Supplementary Figs. and . b Time-dependent changes of the cellular PI content. c Heatmap showing the time-dependent changes of the cellular PI profile ( P = 0.0007). Data are given as percentage of vehicle control for each time point. Mean ( c ) or mean + s.e.m. ( b ) and single data ( a , d – f ) from n = 3 ( a right panel, b , c , e , f ), n = 4 ( a left panel, d ) independent experiments. *** P < 0.001 for the respective time point ( b ) or P values given vs. vehicle control ( a , d – f ); repeated measures one-way ANOVA ( a , d – f ) of log data ( b ) + Tukey HSD post hoc tests.

Article Snippet: CHX, ETO, VAL, I3M, and CAY10566 were purchased from Cayman Chemical (Ann Arbor, MI).

Techniques:

Journal: Nature Communications

Article Title: PI(18:1/18:1) is a SCD1-derived lipokine that limits stress signaling

doi: 10.1038/s41467-022-30374-9

Figure Lengend Snippet: Fibroblasts were cultivated under diverse cytotoxic conditions for 48 h or as indicated. a Negative correlation (−0.6 > r > −1) between cellular p-p38 MAPK (Thr180/Tyr182) levels (at 48 h) and the proportions of phospholipid (PL) species are shown for the co-regulated lipid network described in Supplementary Fig. . Correlations were calculated for mean p-p38 MAPK levels from three independent experiments. b Heatmap showing time-dependent changes in the activation of p38 MAPK compared to vehicle control for each time point. Representative Western blots are shown in Supplementary Fig. . STS, excluded due to pan-kinase inhibition; gray color for I3M, not determined. c , d Phosphorylation and expression of p38 MAPK ( P = 0.99995) ( c ) and JNK ( P = 0.000007) ( d ). Western blots are representative of five ( c ) or three ( b , d ) independent experiments. Data of c is identical to w/o in Supplementary Fig. , and . Mean ( b ) or mean + s.e.m. and single data ( c , d ) from n = 1 ( b I3M for 24 h), n = 2 ( b TNFα for 0.17 h, d for TPG), n = 3 ( a , b , d ), n = 4 ( b , c for CHX, I3M at 48 h), n = 5 ( b for 48 h, c ) independent experiments. P values given vs. vehicle control; mixed-effects model (REML) + Tukey HSD post hoc tests of log data ( c , d ). e Counter-regulation of PI(18:1/18:1) ratios and p38 MAPK activation during VAL-induced cell death across cell lines. MCF-7 breast adenocarcinoma cells, HEK293 embryonic kidney cells, primary human monocytes, MM6 acute monocytic leukemia cells, HT29 colon adenocarcinoma cells, HeLa cervical carcinoma cells, HepG2 hepatoma cells, and HUVECs were treated with vehicle or VAL (10 µM) for 48 h. Percentage changes in cellular PI(18:1/18:1) ratios and p-p38 MAPK levels were calculated vs. vehicle (100%), and the difference to the vehicle control is presented. Representative Western blots are shown in Supplementary Fig. . Detailed descriptions of datasets shown in panel e are given in Supplementary Note . P values given vs. vehicle control; two-tailed paired student t test.

Article Snippet: CHX, ETO, VAL, I3M, and CAY10566 were purchased from Cayman Chemical (Ann Arbor, MI).

Techniques: Activation Assay, Western Blot, Inhibition, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: PI(18:1/18:1) is a SCD1-derived lipokine that limits stress signaling

doi: 10.1038/s41467-022-30374-9

Figure Lengend Snippet: Fibroblasts were cultivated under diverse cytotoxic conditions for 48 h ( a – c , e ) or as indicated ( d ). a Cellular proportion of non-esterified SFAs, MUFAs, and PUFAs. SFAs: 12:0, 14:0, 16:0, 18:0; MUFAs: 16:1, 18:1; PUFAs: 18:2, 20:4, 22:5, 22:6 (MUFA LTR P = 0.0146, 0.0128, 0.0023, 0.0155, 0.003; SFA LTR P = 0.0176, 0.0088, 0.0013, 0.0072, 0.0015). b Heatmap showing changes in the free fatty acid profile as compared to vehicle control. Data are given as percentage of the relative free fatty acid abundance. c Volcano plots highlighting free fatty acids that are strongly and significantly modulated by VAL or MC. Comparisons of the indicated treatment groups show the mean difference of percentage changes and the negative log10(adjusted P value). Adjusted P values given vs. vehicle control; two-tailed multiple unpaired student t tests from log data with correction for multiple comparisons using a two-stage linear step-up procedure by Benjamini, Krieger, and Yekutieli (false discovery rate 5%). d Heatmaps showing the time-dependent effect on Scd1 , Actb , and Gapdh mRNA levels that were normalized to the total amount of cellular RNA and compared to vehicle control for each time point. e Protein expression of SCD1. Western blots are representative of seven independent experiments (LTR P = 0.000000002, \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$0.\bar{99}$$\end{document} 0 . 99 ¯ ). Mean ( b – d ) or mean + s.e.m. ( a ) and single data ( e ) from n = 2 ( d for Scd1 and Gapdh at 6 h; Actb at 48 h for Serum), n = 3 ( a – d ), n = 6 ( e for TNFα, Serum, I3M), n = 7 ( e ) independent experiments. * P < 0.05, ** P < 0.01 or P values given vs. vehicle control; repeated measures one-way ANOVA ( a ) or mixed-effects model (REML) of log data ( e ) + Tukey HSD post hoc tests.

Article Snippet: CHX, ETO, VAL, I3M, and CAY10566 were purchased from Cayman Chemical (Ann Arbor, MI).

Techniques: Two Tailed Test, Expressing, Western Blot