hfd  (PMI Nutrition International LLC)

Journal: Redox Biology

Article Title: Ergothioneine rescues obesity-induced testicular dysfunction via dual restoration of steroidogenesis and mitochondrial redox homeostasis

doi: 10.1016/j.redox.2026.104090

Figure Lengend Snippet: Chronic high-fat diet progressively impairs testicular architecture and sperm quality. (A) Longitudinal body weight changes in control and HFD-fed mice over a 14-week period (left), and representative gross morphology showcasing significant obesity in the HFD group (right). n = 6. (B) Oral glucose tolerance test (OGTT) profiles demonstrating impaired glucose homeostasis in HFD mice. (C) Adipose and organ weights, including epididymal fat, inguinal fat, liver, brown adipose tissue (BAT), and testes, normalized to body weight. (D-E) Quantitative analysis of sperm concentration and total motility assessed by computer-assisted sperm analysis (CASA). (F) Representative H&E-stained sections of testicular tissues from SNC (Short-term Normal Control), SHFD (Short-term HFD), LNC (Long-term Normal Control), and LHFD (Long-term HFD) groups. scale bar = 50 μm. (G-H) Representative immunohistochemical (IHC) staining, morphometric quantification of the Mean Tubular Biopsy Score (MTBS) and germinal epithelium (GE) thickness, reflecting the degree of seminiferous tubule degeneration. (I-J) Quantification of Transition Protein 1 (TNP1) and γH2AX in testicular sections. γH2AX serves as a marker for DNA double-strand breaks in meiotic spermatocytes. scale bar = 20 μm. (K–N) Cytotoxicity profiles of mouse testicular cell lines (GC-1 spg, GC-2 spd, TM3, and TM4) following exposure to varying concentrations of palmitate (PA) for 48 h, assessed via CCK-8 assay. (O) Representative fluorescence images of BODIPY™ 493/503 staining visualizing neutral lipid droplet (LD) accumulation in GC-1 spg, GC-2 spd, TM3, and TM4 cells under lipotoxic conditions. scale bar = 20 μm. Data are presented as mean ± SEM ( n ≥ 3 independent biological replicates). The normality of the data was verified using the Shapiro-Wilk test. Statistical significance was determined by one-way ANOVA followed by Tukey's post-hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 indicate significant differences compared to the control group.

Article Snippet: The HFD + ET group was fed the same diet and, in parallel, administered Ergothioneine (497-30-3; Med Chem Express) by oral gavage at 100 mg/kg/day.

Techniques: Control, Concentration Assay, Staining, Immunohistochemical staining, Immunohistochemistry, Marker, CCK-8 Assay, Fluorescence

Journal: Redox Biology

Article Title: Ergothioneine rescues obesity-induced testicular dysfunction via dual restoration of steroidogenesis and mitochondrial redox homeostasis

doi: 10.1016/j.redox.2026.104090

Figure Lengend Snippet: Progressive depletion of ergothioneine and lipid remodeling defined in the HFD testes . (A) Multivariate partial least squares-discriminant analysis (PLS-DA) score plot illustrating a distinct separation of metabolic phenotypes between the HFD and Control groups at different time points. (B) Volcano plot identifying significantly altered metabolites based on the criteria of variable importance in projection(VIP) > 1.0 and P < 0.05. (C) Chemical class distribution of the identified differential metabolites, highlighting major lipid and organic acid remodeling. (D) Differential abundance of top-ranked testicular metabolites, showing significantly upregulated and downregulated molecules across experimental cohorts. (E) Summary of the total number of differential metabolites identified through pairwise comparisons (SHFD vs. SNC; LHFD vs. LNC; and LHFD vs. SHFD), demonstrating the temporal metabolic shift. (F-G) Venn diagrams depicting intersectional analysis of shared and unique (F) upregulated and (G) downregulated metabolites among the indicated longitudinal comparisons. (H) Longitudinal quantification of l -Ergothioneine (ET) levels in testicular tissues across the SNC, SHFD, LNC, and LHFD groups, showcasing its progressive depletion during HFD-induced obesity. Data are presented as mean ± SEM ( n = 6 biological replicates for metabolomics). The normality of the metabolic data was rigorously assessed using the Shapiro-Wilk test. For multiple group comparisons, one-way ANOVA followed by Tukey's post-hoc test was employed. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus the respective control group.

Article Snippet: The HFD + ET group was fed the same diet and, in parallel, administered Ergothioneine (497-30-3; Med Chem Express) by oral gavage at 100 mg/kg/day.

Techniques: Control

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Grass carp can adapt to short-term HFD by expanding healthy adipose tissue . A , body weight from grass carp–fed Control (normal diet) and HFD (high-fat diet) for 3 days (D3), 2 weeks (W2), 4 weeks (W4), and 8 weeks (W8), n = 3. B , hepatosomatic from grass carp–fed Control and HFD for D3, W2, W4, and W8. Hepatosomatic index (HSI, %) = 100 × (liver weight, g)/(individual fish weight, g), n = 3. C , intraperitoneal fat index from grass carp–fed Control and HFD for D3, W2, W4, and W8. Intraperitoneal fat index (IPFI, %) = 100 × (intraperitoneal fat weight, g)/(final weight, g), n = 3. D , H&E stains of the adipose tissue from grass carp–fed Control and HFD for D3, W2, W4, and W8. Scale bar represents 50 μm. E , using ImageJ software to measure relative adipocyte size combined with H&E staining results of the adipose tissue, n = 3. F , representative images of immunofluorescence staining for EdU in the adipocyte nuclei of adipose tissue from grass carp after 1 week of EdU treatment and 2 weeks on the indicated diet. Tissue is also stained for caveolin-1 ( green ) to visualize adipocyte plasma membranes, as well as DAPI ( blue ) to visualize nuclei. Scale bar represents 50 μm. G , quantitative analysis of EdU positive nuclei in grass carp adipocytes after 3 days, 2 weeks, 4 weeks, and 8 weeks of conditional diet. The yellow arrows indicate edu-positive adipocyte nuclei, which are located within the cytoplasmic membrane of adipocytes, n = 3. H , growth rate of EDU-positive cell nuclei in adipocyte at different time periods, n = 3. I , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. J , quantification of CLS, n = 3. K , the mRNA expression of IL-1β, IL-8, TNF-α in adipose tissue, n = 3. Results are presented as means ± SD. Data analysis was conducted by Student’s two-tailed t test. Results with p < 0.05 represents a statistically significant difference: ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Control, Software, Staining, Immunofluorescence, Clinical Proteomics, Membrane, Immunohistochemical staining, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Short-term (W2) HFD activates the HIF-1 signaling pathway in grass carp adipose tissue and promotes adipose tissue expansion . A , KEGG pathway enrichment bubble plot of the top 10 differentially expressed genes identified by transcriptomic analysis of adipose tissue from grass carp after 2 weeks of Control or HFD feeding, n = 3. B , whole-mount immunohistochemical analysis of adipose tissue of grass carp. Green , pimonidazole adduct; off-white , adipocyte membrane; scale bar represents 100 μm C , the mRNA expression of HIF1αa downstream target genes VEGFα, ANGPTL4, TGFβ in the adipose tissue of grass carp after 2 weeks of Control and HFD feeding, n = 3. D , body weight of grass carp fed with PX-478 added to HFD for 2 weeks, n = 3. E , intraperitoneal fat index of grass carp fed with PX-478 added to HFD for 2 weeks, n = 3. F , triglyceride content in adipose tissue of grass carp fed with PX-478 added to HFD for 2 weeks, n = 3. G , anatomical diagram of abdominal adipose tissue of grass carp, and H&E staining of the adipose tissue from grass carp; scale bar represents 100 μm. H , relative adipocyte size in the adipose tissue of grass carp fed with PX-478 added to HFD for 2 weeks, n = 3. I , quantitative analysis of EDU-positive cell nuclei in the adipose tissue of grass carp fed with PX-478 added to HFD for 2 weeks, n = 3. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Control, Immunohistochemical staining, Membrane, Expressing, Staining, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Short-term (W2) HFD activates HIF1αa-induced ATGL ubiquitination and degradation in mature adipocyte . A , heatmaps of the expression of metabolism and health-related proteins in grass carp adipose tissue following a 2-weeks feeding period with control or high-fat diet, n = 3. B , the mRNA expression of HIF1αa gene in freshly isolated primary mature adipocytes obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. C , the mRNA expression of HIF1αa gene in freshly isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. D , Western blot analysis of ATGL protein levels in isolated primary mature adipocytes obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. D ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. E , Western blot analysis of ATGL protein levels in isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. E ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. F , Western blot analysis of the effect of HIF1αa on the stability of ATGL. HA-ATGL and EGFP-HIF1αa plasmids were cotransfected into 293T cells and treated with MG132 (10 μM) 6 h before termination. F ′, quantitative analysis of immunoblots was performed, n = 3. G , Western blot analysis of the effect of HIF1αa on the ubiquitination of ATGL. 293T cells were transfected with the indicated plasmids for 24 h, for the last 6 h in the presence of MG132 (10 μM). Data are represented as the mean ± SD. Statistical testing was performed with one-way ANOVA. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Ubiquitin Proteomics, Expressing, Control, Isolation, Western Blot, Transfection

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: HIF1αa promotes adipocyte hypertrophy of grass carp by degrading ATGL through the ubiquitinated proteasome pathway . A , treat differentiated adipocytes (day 8) with 350 μM CoCl 2 for 12, 24, and 48 h and detect their ATGL protein expression levels. β-actin protein was used as a loading control. A ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. B , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 12 h and perform BODIPY staining. C , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 12 h and perform triglyceride content detection, n = 3. D , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 12 h and detect their ATGL protein expression levels. β-actin protein was used as a loading control. D ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. E , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 12 h and detect the ubiquitination level of ATGL by IP. MG132 (10 μM) was added 6 h before sample collection. F , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 48 h and perform BODIPY staining. G , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 48 h and perform triglyceride content detection, n = 3. H , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 48 h and detect their ATGL protein expression levels. β-actin protein was used as a loading control. H ′, quantitative analysis of immunoblots was performed, n = =2 independent experiments. I , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 48 h and detect the ubiquitination level of ATGL by IP. MG132 (10 μM) was added 6 h before sample collection. Data are represented as the mean ± SD. Statistical testing was performed with one-way ANOVA. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Expressing, Control, Western Blot, Staining, Ubiquitin Proteomics

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: HIF1αa promotes ATGL mRNA expression through transcriptional regulation, maintaining basal lipolysis in mature adipocyte . A , treat differentiated adipocytes with 350 μM CoCl 2 for 12, 24, and 48 h and detect the expression levels of HIF1αa and ATGL mRNA in the cells, n = 3. B , treat differentiated adipocytes with 350 μM CoCl 2 for 48 h and collect the culture medium to detect free fatty acid (FFA) content, n = 3. C , ex vivo FFA release by mature adipocytes isolated from HFD-fed grass carp for 2 weeks, n = 3. D , site-mutation analysis of ATGL-binding sites on pGL3.0-HIF1αa vectors in 293 T cells, n = 3. E , treat differentiated adipocytes with 350 μM CoCl 2 and PX-478 (10 μM) for 48 h and detect the expression levels of HIF1αa and ATGL mRNA in the cells, n = 3. F , treat differentiated adipocytes with CoCl 2 (350 μM) and PX-478 (10 μM) for 48 h and collect the culture medium to detect free fatty acid (FFA) content, n = 3. G , the mRNA expression of ATGL gene in freshly isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. H , the mRNA expression of ATGL gene in freshly isolated primary mature adipocytes obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Expressing, Ex Vivo, Isolation, Mutagenesis, Binding Assay, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Inhibition of adipocyte hyperplasia and induction of pathological expansion of adipose tissue under short-term HFD conditions . A , body weight of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. B , intraperitoneal fat index of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. C , anatomical diagram of abdominal adipose tissue of grass carp and its H&E staining; scale bar represents 50 μm. D , triglyceride content in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. E , relative adipocyte size in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. F , relative adipocyte number in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. G , quantitative analysis of EDU-positive cell nuclei in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. H , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. H ′, quantification of CLS, n = 3. I , the mRNA expression of IL-1β, IL-8 in adipose tissue, n = 3. J , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, HFD + PX-478, and HFD + GW9662. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. J ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. K , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 3 days of Control and HFD. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. K ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Inhibition, Staining, Immunohistochemical staining, Expressing, Western Blot, Isolation, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Grass carp can adapt to short-term HFD by expanding healthy adipose tissue . A , body weight from grass carp–fed Control (normal diet) and HFD (high-fat diet) for 3 days (D3), 2 weeks (W2), 4 weeks (W4), and 8 weeks (W8), n = 3. B , hepatosomatic from grass carp–fed Control and HFD for D3, W2, W4, and W8. Hepatosomatic index (HSI, %) = 100 × (liver weight, g)/(individual fish weight, g), n = 3. C , intraperitoneal fat index from grass carp–fed Control and HFD for D3, W2, W4, and W8. Intraperitoneal fat index (IPFI, %) = 100 × (intraperitoneal fat weight, g)/(final weight, g), n = 3. D , H&E stains of the adipose tissue from grass carp–fed Control and HFD for D3, W2, W4, and W8. Scale bar represents 50 μm. E , using ImageJ software to measure relative adipocyte size combined with H&E staining results of the adipose tissue, n = 3. F , representative images of immunofluorescence staining for EdU in the adipocyte nuclei of adipose tissue from grass carp after 1 week of EdU treatment and 2 weeks on the indicated diet. Tissue is also stained for caveolin-1 ( green ) to visualize adipocyte plasma membranes, as well as DAPI ( blue ) to visualize nuclei. Scale bar represents 50 μm. G , quantitative analysis of EdU positive nuclei in grass carp adipocytes after 3 days, 2 weeks, 4 weeks, and 8 weeks of conditional diet. The yellow arrows indicate edu-positive adipocyte nuclei, which are located within the cytoplasmic membrane of adipocytes, n = 3. H , growth rate of EDU-positive cell nuclei in adipocyte at different time periods, n = 3. I , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. J , quantification of CLS, n = 3. K , the mRNA expression of IL-1β, IL-8, TNF-α in adipose tissue, n = 3. Results are presented as means ± SD. Data analysis was conducted by Student’s two-tailed t test. Results with p < 0.05 represents a statistically significant difference: ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Control, Software, Staining, Immunofluorescence, Clinical Proteomics, Membrane, Immunohistochemical staining, Expressing, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Short-term (W2) HFD activates HIF1αa-induced ATGL ubiquitination and degradation in mature adipocyte . A , heatmaps of the expression of metabolism and health-related proteins in grass carp adipose tissue following a 2-weeks feeding period with control or high-fat diet, n = 3. B , the mRNA expression of HIF1αa gene in freshly isolated primary mature adipocytes obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. C , the mRNA expression of HIF1αa gene in freshly isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478, n = 3. Each sample is composed of 20 fish collected from the same experimental group and tank. D , Western blot analysis of ATGL protein levels in isolated primary mature adipocytes obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. D ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. E , Western blot analysis of ATGL protein levels in isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, and HFD + PX-478. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. E ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. F , Western blot analysis of the effect of HIF1αa on the stability of ATGL. HA-ATGL and EGFP-HIF1αa plasmids were cotransfected into 293T cells and treated with MG132 (10 μM) 6 h before termination. F ′, quantitative analysis of immunoblots was performed, n = 3. G , Western blot analysis of the effect of HIF1αa on the ubiquitination of ATGL. 293T cells were transfected with the indicated plasmids for 24 h, for the last 6 h in the presence of MG132 (10 μM). Data are represented as the mean ± SD. Statistical testing was performed with one-way ANOVA. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Ubiquitin Proteomics, Expressing, Control, Isolation, Western Blot, Transfection

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: HIF1αa induces the release of FFA from mature adipocytes through basal lipolysis, which activates PPARγ in preadipocytes, promoting adipocyte hyperplasia . A , schematic drawing of the experimental procedures. B , treat differentiated adipocytes with 350 μM CoCl 2 for 48 h; the growth medium was replaced to continue culturing the mature adipocytes for an additional 48 h, and the medium was collected to measure the free fatty acid (FFA) content, n = 3. C , BODIPY staining of grass carp primary preadipocytes under different medium treatment conditions, Bodipy ( green ) and nuclei ( blue ). D , differential genes related to adipogenesis, lipid metabolism, inflammation, and fibrosis in transcriptome sequencing of preadipocytes under different culture media treatment conditions, n = 3. E , transcriptional activity of PPAR γ under different culture media and OA treatment conditions, n = 3. F , the differentiated adipocytes were treated with 350 μM CoCl 2 and 1 μM BODIPY 558/568 C 12 (a fluorescent fatty acid tracer) ( red ) for 48 h to induce lipolysis and FFA labeling. After washing to remove residual compounds, differentiated adipocytes were cocultured with preadipocytes for 48 h. Use Bodipy ( green ) staining and take photos with a confocal laser microscope. Control group: preadipocytes cocultured with differentiated adipocytes that were not treated with CoCl 2 ; CoCl 2 group: preadipocytes cocultured with differentiated adipocytes that were treated with CoCl 2 ; CoCl 2 + GW9662 group: preadipocytes cocultured with differentiated adipocytes that were treated with CoCl 2 and then treated with GW9662. G , TG content of preadipocytes under different coculture systems and composite treatment conditions, n = 3. H , the mRNA expression levels of PPARγ and DGAT2 in preadipocytes under different coculture systems and composite treatment conditions, n = 3. I , Western blot of PPARγ in preadipocytes under different coculture systems and composite treatment conditions. β-actin protein was used as a loading control. I ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Staining, Sequencing, Activity Assay, Labeling, Microscopy, Control, Expressing, Western Blot, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Hypoxia-inducible factor 1αa regulates lipid metabolism to coordinate adipocyte hypertrophy and hyperplasia in grass carp

doi: 10.1016/j.jbc.2026.111195

Figure Lengend Snippet: Inhibition of adipocyte hyperplasia and induction of pathological expansion of adipose tissue under short-term HFD conditions . A , body weight of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. B , intraperitoneal fat index of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. C , anatomical diagram of abdominal adipose tissue of grass carp and its H&E staining; scale bar represents 50 μm. D , triglyceride content in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. E , relative adipocyte size in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. F , relative adipocyte number in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. G , quantitative analysis of EDU-positive cell nuclei in the adipose tissue of grass carp fed with GW9662 added to HFD for 2 weeks, n = 3. H , immunohistochemical staining with an F4/80 antibody of adipose tissue, crown-like structures (CLS); scale bar represents 50 μm. H ′, quantification of CLS, n = 3. I , the mRNA expression of IL-1β, IL-8 in adipose tissue, n = 3. J , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 2 weeks of Control, HFD, HFD + PX-478, and HFD + GW9662. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. J ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. K , Western blot analysis of PPARγ protein levels in isolated SVF obtained directly from grass carp adipose tissue after 3 days of Control and HFD. Each sample is composed of 20 fish collected from the same experimental group and tank. β-actin protein was used as a loading control. K ′, quantitative analysis of immunoblots was performed, n = 2 independent experiments. Data are represented as the mean ± SD. For comparisons between two groups, a two-tailed Student’s t test was performed, ∗ p < 0.05. Statistical testing was performed with one-way ANOVA for comparisons among three or more groups. The significance of the differences was assessed with post hoc Tukey test. Set at p < 0.05 shows the significant level. Different lowercase letter means significant difference among groups.

Article Snippet: Five experimental diets were prepared in this experiment: Control group (protein 33%, lipid 4%), HFD group (protein 33%, lipid 8%), HFD + PX-478 (HY-10231, MCE) group (protein 33%, lipid 8% + PX-478 60 mg/kg), and HFD + GW9662 (HY-16578, MCE) group (protein 33%, lipid 8% + GW9662 60 mg/kg).

Techniques: Inhibition, Staining, Immunohistochemical staining, Expressing, Western Blot, Isolation, Control, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a – c A transcriptome analysis of mature adipocytes isolated from EpiWAT of C57BL/6J mice on 16-week NCD and HFD. a Network diagram illustrating the interaction among the top 10 immune-related pathways enriched by KEGG analysis of DEGs. The table showing the degree of interaction for each pathway, as assessed by the cumulative interaction scores of the genes within the pathway. b GSEA of DEGs showing two B2M-associated pathways in adipocytes. c Network diagram illustrating the interaction among genes in antigen processing and presentation pathway (blue) and iron uptake and transport pathway (green), with B2M (red) at the intersection. Relative B2m mRNA levels ( d ), protein expressions ( e ) and relative quantification ( f ) in EpiWAT, SVF and purified mature adipocyte fraction from EpiWAT of 16-week NCD- or HFD-fed mice ( n = 3-8). g Representative fluorescence images of B2M (green) and DAPI (blue) staining in EpiWAT sections of NCD-fed (upper) and HFD-fed (below) mice. The right panel quantifies the area-normalized B2M MFI (n = 3). Scale bar = 50 μm. Relative B2m mRNA levels ( h ), protein expressions ( i ) and relative quantification ( j ) in 3T3-L1 adipocytes treated with PA at the indicated dose for 24 h ( n = 4-5). The data are representative as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significant in d , f , and g was calculated using a two-tailed Student’s t test. Significant in h and j was calculated using one-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001. B2M β2-microglobulin, DEGs differentially expressed genes, EpiWAT epididymal adipose tissue, HFD high-fat diet, MFI mean fluorescence intensity, NCD normal chow diet, PA palmitic acid, SVF stromal vascular fraction

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Isolation, Quantitative Proteomics, Purification, Fluorescence, Staining, Standard Deviation, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a Study design of diet induced obesity model. b , c Representative photograph of B2m f/f and B2m cKO mice and their respective fat pads. d Body weights ( n = 8). e Weight and proportion of EpiWAT ( n = 8). f Body content (fat, lean, water) analyzed by NMR spectrometer ( n = 6–8). Representative histological images showing H&E staining and Oil Red O staining of indicated sections of HFD-fed B2m f/f and B2m cKO mice ( g ), with quantitation of average adipocyte size of EpiWAT and SAT section ( h ), NAS ( i ), and quantitation of Oil Red O staining ( j ) ( n = 3). Scale bars, 100 μm. k – n Serum levels of leptin, adiponectin, insulin, and glucagon in HFD-fed B2m f/f and B2m cKO mice after 4 hours of fasting ( n = 6). o , p IPGTT (2 g glucose per kg body weight), IPITT (0.75 U insulin per kg body weight) and their respective AUC ( n = 8). q – t Serum levels of TG, TC, HDL and LDL ( n = 8). u Serum levels of TNFα, IFNγ, IL-1β, and IL-6 ( n = 7-8). The data are represented as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significance in e , f , h , j – n , AUC in o , AUC in p , and q – u was calculated using a two-tailed Student’s t test. Significance in d , o , and p was calculated using two-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. Significance in i was calculated using the Mann-Whitney U test. * p < 0.05, ** p < 0.01, *** p < 0.001. AUC area under the curve, BAT brown adipose tissue, EpiWAT epididymal adipose tissue, HDL high density lipoprotein, HFD high-fat diet, IPGTT intraperitoneal glucose tolerance test, IPITT intraperitoneal insulin tolerance test, LDL low density lipoprotein, NAS Non-alcoholic fatty liver disease activity score, NCD normal chow diet, SAT subcutaneous adipose tissue, TC cholesterol, TG triglyceride

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Staining, Quantitation Assay, Standard Deviation, Two Tailed Test, MANN-WHITNEY, Activity Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a GSEA of antigen processing and presentation pathway between HFD-fed WT mice and B2m cKO mice. Relative mRNA levels of H2-K1 ( b ) and H2-D1 ( c ) in isolated adipocyte fraction of EpiWAT from NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 4). Protein expressions and relative quantitation of MHC Class I in whole cell lysates ( d , e ) and membrane component ( f , g ) derived from adipocyte fraction isolated from EpiWAT of NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 3). h Schematic representation of the in vitro co-culture experiment involving primary adipocytes and CD8 + T cells. CD8 + T cells were isolated from EpiWAT of obese B2m f/f mice using a magnetic bead sorting kit and labeled with CFSE before co-culture. Adipocytes were induced from primary adipocyte precursors in SVF and treated with or without PA (2.5 mM) for 24 hours before co-culture. Flow cytometric analysis of proliferation ( i , j ) and CD69 ( k , l ) expression in CD8 + T cells following co-culture with primary adipocytes. ( n = 3). The data are representative as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significant in b – d , and f was calculated using two-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. Significant in j and l was calculated using multi-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001. CFSE Carboxyfluorescein Succinimidyl Ester, EpiWAT epididymal adipose tissue, HFD high fat diet, NCD normal chow diet, PA palmitic acid, SVF stromal vascular fraction, WT wild type

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Isolation, Quantitation Assay, Membrane, Derivative Assay, In Vitro, Co-Culture Assay, Labeling, Expressing, Standard Deviation

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a Relative mRNA levels of Fth and Ftl in the adipocyte fraction from EpiWAT of NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 8). Protein expressions ( b ) and relative quantitation normalized to the internal reference Actin ( c ) of FTH1 and FTL1 in the lysates of mature adipocytes isolated from EpiWAT of NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 3). d Total iron content, as measured by iron assay kit, in mature adipocytes from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 8). e The correlation of total iron content in ( d ) with EpiWAT proportion (%) ( n = 32). f Co-immunoprecipitation assay using HFE as bait protein showing the interaction between HFE, TFR1, and TFR2 within the membrane fractions of adipocytes isolated from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice. Relative mRNA levels of Tfr2 ( g ), Hfe ( h ), Tfr1 ( i ), and HAMP ( j ) in the adipocyte fraction isolated from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 7-8). Protein expressions ( k ) and relative quantitation normalized to the internal reference Vinculin in TFR2 ( l ), HFE ( m ), TFR1 ( n ), hepcidin ( o ), and FPN ( p ) in the lysates of adipocyte fraction isolated from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 3). Protein expressions ( q ) and relative quantitation normalized to the internal reference Na,K-ATPase of TFR2 ( r ), HFE ( s ), TFR1 ( t ), and FPN ( u ) in membrane component derived from adipocyte fraction isolated from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 3). The data are represented as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significant in a , c , d , g – j , l – p , and r – u was calculated using two-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001.EpiWAT epididymal adipose tissue, FTH ferritin heavy chain, FTL ferritin light chain, HFD high fat diet, HFE Hereditary hemochromatosis protein, NCD normal chow diet, TFR1 transferrin receptor 1, TFR2 transferrin receptor 2, FPN ferroportin

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Quantitation Assay, Isolation, Iron Assay, Co-Immunoprecipitation Assay, Membrane, Derivative Assay, Standard Deviation

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a – g Adipocytes were induced from primary adipocyte precursors in SVF of NCD-fed B2m f/f and B2m cKO mice and treated with or without PA (2.5 mM) for 24 hours. a cell viability ( n = 4). The levels of ROS ( b , c ), lipid peroxides ( d , e ), and Fe 2+ ( f , g ) in adipocytes were assessed using DCFH, C11-BODIPY 581/591 , and FeRhoNox-1 probes, respectively ( n = 3-6). h – m Adipocytes were induced from primary adipocyte precursors in SVF from EpiWAT of NCD-fed B2m f/f and B2m cKO mice and treated with or without PA (2.5 mM) and/or Erastin (2 μM) for 24 hours. The levels of ROS ( h , i ), lipid peroxides ( j , k ), and Fe 2+ ( l , m ) in adipocytes were assessed using DCFH, C11-BODIPY 581/591 , and FeRhoNox-1 probes, respectively ( n = 3-5). n Ferrous ion content in mature adipocytes from NCD- or HFD-fed B2m f/f and B2m cKO mice, measured using an iron content detection kit ( n = 8). o The correlation of ferrous ion content in ( n ) with EpiWAT proportion (%) in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 32). Representative images ( p ) and relative quantification ( q ) of ROS, visualized using staining fluorescent probe DHE, in EpiWAT of HFD-fed B2m f/f and B2m cKO mice, DAPI (blue). ( n = 5). Scale bar, 100 μm. Levels of MDA ( r ) and GSH ( s ) in mature adipocytes isolated from EpiWAT in NCD- or HFD-fed B2m f/f and B2m cKO mice ( n = 8). t Representative transmission electron microscopy images of the mitochondria (indicated by red arrow) of adipocytes in EpiWAT of NCD- or HFD-fed B2m f/f and B2m cKO mice. ( n = 5-6). u Flow cytometry analysis of CD86, CD80, and CD11c expressions on ATMs following 24 hours of co-culture with primary adipocytes. ATMs were isolated from EpiWAT of lean B2m f/f mice via F4/80 + magnetic beads. Primary adipocytes were differentiated from adipocyte precursors in SVF from EpiWAT of lean B2m f/f or B2m cKO mice and treated with or without PA (2.5 mM) for 24 hours prior to co-culture ( n = 4). v Representative immunofluorescence staining images of ATMs in EpiWAT of NCD- or HFD-fed B2m f/f and B2m cKO mice, showing perilipin (green), 4-HNE (orange), F4/80 (cyan), CD206 (yellow), CD11c (red), and DAPI (blue). Scale bar, 50 μm. The data are represented as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significance in q was calculated using a two-tailed Student’s t test. Significant in a , c , e , g , i , k , m , n , r , s , and u was calculated using two-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001. 4-HNE 4-hydroxynonenal, ATMs adipose tissue macrophages, DHE Dihydroethidium, EpiWAT epididymal adipose tissue, GSH glutathione, HFD high fat diet, MDA malondialdehyde, NCD normal chow diet, PA palmitic acid, ROS reactive oxygen species, SVF stromal vascular fraction

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Quantitative Proteomics, Staining, Isolation, Transmission Assay, Electron Microscopy, Flow Cytometry, Co-Culture Assay, Magnetic Beads, Immunofluorescence, Standard Deviation, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: a Overview for AAV9-mediated knockdown of B2M in EpiWAT. b Experiment scheme of male mice was injected with either AAV9-Fabp5-ZsGreen- B2m- shRNA (AAV9- B2m ) or AAV9-Fabp5-ZsGreen (AAV9-null) following six-week HFD feeding and subsequently maintained on the same diet for an additional 6 weeks. c Ex vivo organ fluorescence imaging of EpiWAT, Heart, Liver, Spleen, Lung and Kidney from AAV9- B2m mice and age-matched sham-operated mice at 2 weeks post-injection. d Fluorescence intensity of tissues in c ( n = 3). e Relative B2m mRNA expression in EpiWAT, Heart, Liver, Spleen, Lung and Kidney ( n = 3). Protein expression ( f ) and relative quantification ( g ) of B2M in adipocytes derived from EpiWAT ( n = 3). h Representative photograph of AAV9-null and AAV9- B2m mice. i Body weights ( n = 6). j Weight and proportion of EpiWAT ( n = 6). Representative histological images showing H&E staining of EpiWAT ( k ), with quantitation of average adipocyte size ( l ) ( n = 3). Scale bar, 100 μm. m – o Representative histological images showing H&E staining and Oil Red O staining of liver sections, with NAS ( n ) and relative quantitation of Oil Red O staining ( o ) ( n = 3). Scale bar, 100 μm. p – s IPGTT (2 g glucose per kg body weight), IPITT (0.75 U insulin per kg body weight) and their respective AUC ( n = 6). t Serum levels of TG, TC, HDL and LDL ( n = 3). Relative mRNA levels of inflammatory cytokines ( u ) and adipocytokines ( v ) in EpiWAT ( n = 3). The data are representative as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significant in d , e , g , j , l , o , q , and s – v was calculated using a two-tailed Student’s t test. Significant in i, p and r was calculated using two-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. Significant in n was calculated using Mann-Whitney U test. * p < 0.05, ** p < 0.01, *** p < 0.001. AAV9 type 9 adeno-associated virus, AUC area under the curve, EpiWAT epididymal adipose tissue, HDL high density lipoprotein, HFD high-fat diet, IPGTT intraperitoneal glucose tolerance test, IPITT intraperitoneal insulin tolerance test, LDL low density lipoprotein, NAS Non-alcoholic fatty liver disease activity score, Sham sham-operated group, TC cholesterol, TG triglyceride

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Knockdown, Injection, shRNA, Ex Vivo, Fluorescence, Imaging, Expressing, Quantitative Proteomics, Derivative Assay, Staining, Quantitation Assay, Standard Deviation, Two Tailed Test, MANN-WHITNEY, Virus, Activity Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: KEGG bubble diagram ( a ) and GO bar chart ( b ) displaying selected enrichment pathways of DEGs in SAT from lean and metabolically unhealthy obese patients. c Relative expression of B2m , HLA-A , HLA-B , HLA-C , Fth and Ftl gene in SAT across lean patients ( n = 11), metabolically healthy obese patients ( n = 14) and metabolically unhealthy obese patients ( n = 20). d Correlation analysis of indicated genes and the Mantel test analysis of clinical indicators (BMI, HOMA-IR and mean adipocyte size) and indicated genes among lean and obese-unhealthy patients. A color gradient is utilized to denote Person’s correlation with the value reflecting the precise value of this coefficient. The color of the line indicates the correlation direction in the Mantel test (Mantel’s r. sign), with red indicating a positive correlation and blue signifying a negative correlation. The thickness of the lines reflects the strength of the correlation in the Mantel test (Mantel’s r.abs): thin lines for weak (r < 0.1), standard lines for moderate (0.1 < r < 0.3), and thick lines for strong (r >= 0.3) correlations. The type of line denotes the significance of the Mantel test (Mantel’s p), with solid indicating a significant correlation and dashed signifying a non-significant correlation. e GSEA of DEGs showing significant upregulation of three pathways in SAT from lean patients and metabolically unhealthy obese patients. f Relative expression of B2m in VAT. GSE286454 dataset with lean (n = 4) and obese (n = 3) and GSE283367 ) with lean (n = 70) and obese (n = 43). g – j Protein expressions and relative quantification of B2M, MHC Class I, TFR2, TFR1, FPN, HFE, FTH, FTL and hepcidin in whole cell lysates derived from adipocyte fraction isolated from VAT across lean patients and obese patients ( n = 3). The data are representative as the mean ± standard deviation (SD), with “n” representing the number of biological replicates per experimental group. Significant in f , h , and j was calculated using a two-tailed Student’s t test. Significant in c was calculated using one-way ANOVA followed by Tukey’s HSD post hoc test for multiple comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001. B2M β2-microglobulin, BMI body mass index, DEGs differentially expressed genes, FTH ferritin heavy chain, FTL ferritin light chain, HFE Hereditary hemochromatosis protein, HOMA-IR homeostasis model assessment of insulin resistance, SAT subcutaneous adipose tissue, TFR1 transferrin receptor 1, TFR2 transferrin receptor 2, FPN ferroportin, VAT visceral adipose tissue

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Metabolic Labelling, Expressing, Quantitative Proteomics, Derivative Assay, Isolation, Standard Deviation, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Adipocytes orchestrate obesity-related chronic inflammation through β2-microglobulin

doi: 10.1038/s41392-025-02486-3

Figure Lengend Snippet: A schematic illustration of the proposed mechanism by which hypertrophic adipocytes induce chronic adipose inflammation and metabolic disorders associated with obesity. Excessive energy stimulates adipocytes to upregulate B2M expression, which not only facilitates the activation and proliferation of CD8 + T cells in adipose tissue through the endogenous antigen presenting pathway, but also promotes iron overload and subsequent ferroptosis of hypertrophic adipocytes via the HFE/B2M-TFR2-Hepcidin-FPN axis, leading to the polarization of ATMs towards M1, thereby accelerating chronic inflammation metabolic disorders. ATMs, adipose tissue macrophages. This figure was created using Adobe Illustrator

Article Snippet: CD8 + T cells were isolated from the EpiWAT of HFD-fed B2m f/f mice via magnetic beads (130-104-075, Miltenyi Biotec).

Techniques: Expressing, Activation Assay