Journal: The FASEB Journal

Article Title: Lymphocyte Antigen 6G Mediates Vagotomy‐Associated Reduction in Body Weight

doi: 10.1096/fj.202600151RR

Figure Lengend Snippet: Vagotomy promoted Ly6G + cell infiltration into eWAT. Wild‐type mice were subjected to left cervical vagotomy (VX) or sham surgery and tissues were collected at 7 days following surgery. (A) The percentage of non‐adipocyte nuclei of total nuclei per section was quantified using ImageJ ( n = 4) and right panels show representative images of paraffin sections of eWAT stained with H&E in sham and VX animals. (B) CCL2 release from eWAT was analyzed by ELISA. The bar shows the CCL2 levels from sham ( n = 4) or VX ( n = 4) mice normalized to eWAT weight: ng/mL per g ± SEM (unpaired Student's t test). (C) eWAT was collected at 1 ( n = 3), 4 ( n = 4 sham, n = 5 VX), and 7 ( n = 15) days following VX or sham surgery and the eWAT SVCs were analyzed by flow cytometry. The bar shows the % ± SEM of CD11b + Ly6G + cells from CD45 + (one‐way ANOVA, Uncorrected Fisher's LSD). (D) Graphs show representative gating for CD11b + Ly6G + cells in sham and VX eWAT at 7 days (concatenated n = 5–6). (E) Representative immunostaining of Ly6G (red) and Perilipin1 (green) in paraffin sections of eWAT. (F–H) Bone marrow neutrophils after sham ( n = 9) or VX ( n = 5) surgery were isolated using negative magnetic beads and analyzed using bulk RNAseq (DESeq2). Heatmap (F), volcano plot (G) of differentially expressed genes, and GO (Gene Ontology) (H) enrichment bar plot. ns = not significant, * p < 0.05. VX, Vagotomy; eWAT, epididymal white adipose tissue; H&E, hematoxylin–eosin; SVCs, stromal vascular cells.

Article Snippet: In vitro release rate was calculated as mg NEFAs per mg eWAT tissue per hour. (2) CCL2 levels were quantified using a mouse CCL2 ELISA kit (R&D Systems, #DY497‐05), according to the manufacturer's instructions ( n = 1 experiment).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Immunostaining, Isolation, Magnetic Beads, RNA sequencing

Journal: Bone Research

Article Title: Expansion of bone marrow adipocytes in obese mice leads to PD-L1-driven bone marrow immunosuppression and osteoclastogenesis

doi: 10.1038/s41413-026-00509-5

Figure Lengend Snippet: Obesity parameters and metabolic phenotype in male B6 mice. a Body weight (g) measurements over 12 weeks for LFD and HFD-fed male B6 mice ( n = 32-41). b Change in weight (%) (dotted line at 28% represents parameter for LFD and dotted line at 58% represents parameter for HFD). c Fold change in fat mass (g) (dotted line at 1.0 represents parameter for LFD and dotted line at 3.3 represents parameter for obese-HFD). d Pearson correlation analysis showing that the change in weight or fat mass fold change both negatively correlate to trabecular bone loss. e Correlation matrix that shows the change in weight or fat mass fold change both negatively correlate to trabecular bone loss (vertical line and horizontal line represent obesity cutoff). f Glucose tolerance test (GTT) for LFD ( n = 26), obese HFD-fed (OB-HFD; n = 34) and non-obese HFD-fed (NO-HFD; n = 7). g Insulin tolerance test (ITT) for LFD, obese HFD-fed (OB-HFD), and non-obese HFD-fed (NO-HFD). h Serum adiponectin levels ( n = 7). i Serum leptin levels. j Serum procollagen type I N-propeptide (P1NP) levels. k Serum tartrate-resistant acid phosphatase 5b (TRAcP 5b) levels. Analyses for a , f , and g were performed as 2-way ANOVA with Šídák’s multiple comparisons test. Significance for the post-hoc analysis for f , g was defined as: * P < 0.05 LFD vs. OB-HFD, # P < 0.05 LFD vs. NO-HFD-fed, and $ P < 0.05 OB-HFD vs. NO - HFD-fed. Analyses for h , i and k were performed as a Kruskal-Wallis test with Dunn’s multiple comparison. Analysis for j was performed as a One-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: Serum protein analysis of adiponectin (Mouse Adiponectin/Acrp30 Quantikine ELISA, R&D Systems MRP300), leptin (Mouse/Rat Leptin Quantikine ELISA, R&D Systems MOB00B), TRAP (Mousetrap TRAcP 5b ELISA, Immunodiagnostic Systems SB-TR103), CTX-1 (Mouse CTX ELISA Kit, Immunodiagnostic Systems AC-06F1), and P1NP (Rat/Mouse P1NP EIA, Immunodiagnostic Systems AC-33F1) were measured by ELISA according to manufacturer’s instructions.

Techniques: Comparison

Journal: The Journal of Clinical Investigation

Article Title: Characterization of intestinal immune responses in generalized human and murine lipodystrophy

doi: 10.1172/JCI192322

Figure Lengend Snippet: ( A ) Representative images of Pparg fl/fl Adipoq-Cre (Lipo) and WT mice showing the complete absence of fat tissue and H&E-stained images of mesenteric adipose tissue (arrows indicate mesenteric and gonadal fat tissue). Scale bars: 100 μm. ( B ) Leptin plasma levels (WT n = 4; Lipo n = 4). ( C ) Liver weights of WT mice ( n = 16) and Pparg fl/fl Adipoq-Cre mice ( n = 12) from 2 independent experiments with a representative image. ( D – G ) Flow cytometric assessment of splenic NK cells from Pparg fl/fl Adipoq-Cre mice ( n = 9–24) compared with WT littermate cells ( n = 10–23). Data were pooled from 3 independent experiments. ( H ) Representative H&E-stained images and immune cell counts (millions/cm) in the terminal ileum or colon as assessed by histology (WT: n = 12–16; Lipo: n = 8–12). Scale bars: 100 μm. ( I ) Gating strategy of T cells and CD4 + FoxP3 + Tregs. The line in the box indicates the median. Boxes range from the 25th to 75th percentiles. Whisker plots show the minimum (smallest) and maximum (largest) values while the line in the box indicates the median. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, by unpaired 2-tailed t test with Welch’s correction.

Article Snippet: Plasma levels of leptin or anti– E. coli (LPS) IgG were determined using the mouse Leptin DuoSet ELISA kit (R&D Systems) or the mouse anti– E. coli LPS (O111:B4) Antibody ELISA Kit (Chondrex).

Techniques: Staining, Clinical Proteomics, Whisker Assay

Journal: The Journal of Clinical Investigation

Article Title: Characterization of intestinal immune responses in generalized human and murine lipodystrophy

doi: 10.1172/JCI192322

Figure Lengend Snippet: ( A ) Experimental design: After 1 DSS cycle (1.5%), lipodystrophic or WT mice were transplanted with 500–600 mg adipose tissue from WT or leptin-deficient ob/ob donors by mini-laparotomy before undergoing another 2 cycles of DSS. Image on right shows vascularized transplanted fat 1 month after surgery. ( B ) Weight change of transplanted fat tissue relative to baseline. ( C ) Plasma leptin levels ( n = 4–9). ( D ) Representative images of H&E- stained colon sections from transplanted versus nontransplanted DSS-treated animals. Scale bars: 100 μm. ( E ) Box-and-whisker plots summarizing the histologic inflammation score of fat-transplanted and nontransplanted animals. Data shown were pooled from 2 independent transplantation experiments (bold symbols) and additional control data points derived from nontransplantation DSS experiments (light gray) shown in ( n = 4–18). ( F ) Liver weights of transplanted WT and Pparg fl/fl Adipoq-Cre mice ( n = 4–18; data were pooled from 5 experiments). ( G ) Representative FACS plots showing IFN-γ and IL-17A production in colonic CD4 + T cells. UNSTIM., unstimulated. ( H ) Box-and-whisker plots summarizing absolute numbers of IFN-γ– and IL-17A–producing CD4 + T cells normalized to WT mice ( n = 4–18; data were pooled from 5 experiments). Statistical differences were calculated by 1-way ANOVA with Šídák’s correction. Each point represents 1 mouse; boxes range from the 25th-75th percentiles. Whisker plots show the minimum (smallest) and maximum (largest) values while the line in the box indicates the median. ( I ) Experimental setup and representative FACS plots showing IFN-γ– and IL-17A–producing CD4 + T cells in peripheral blood of a patient with AGLCD before and 4 days after daily recombinant leptin substitution. transpl., transplantation. Data indicate the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, by 1-way ANOVA with Tukey’s multiple comparisons test for B , C , E , F , and H .

Article Snippet: Plasma levels of leptin or anti– E. coli (LPS) IgG were determined using the mouse Leptin DuoSet ELISA kit (R&D Systems) or the mouse anti– E. coli LPS (O111:B4) Antibody ELISA Kit (Chondrex).

Techniques: Clinical Proteomics, Staining, Whisker Assay, Transplantation Assay, Control, Derivative Assay, Recombinant