recombinant mouse leptin  (R&D Systems)

Journal: Frontiers in Nutrition

Article Title: Astragaloside IV modulates oxidative stress and osteoimmune–Wnt signaling in ovariectomized rats: an integrated study of RNA sequencing, molecular docking, and experimental validation

doi: 10.3389/fnut.2026.1785452

Figure Lengend Snippet: Molecular docking models of Astragaloside IV (AS-IV) binding to the core targets. Representative binding poses of AS-IV (shown in stick representation) within the predicted binding pockets of the key target proteins. The calculated binding free energy (kcal/mol) for each complex is indicated. (A) Il22ra2 (−10.2 kcal/mol), (B) Ptgs2 (−9.5 kcal/mol), (C) Wnt1 (−9.3 kcal/mol), (D) Igfbp2 (−8.4 kcal/mol), (E) GZMB (−8.2 kcal/mol), (F) Lep (−7.8 kcal/mol), (G) Gfap (−7.8 kcal/mol), (H) Wnt10a (−7.7 kcal/mol). These results demonstrate stable binding interactions between the therapeutic compound AS-IV and all eight core targets identified from the PPI network and WGCNA, with Il22ra2 and Ptgs2 showing the strongest binding affinity.

Article Snippet: Primary antibodies diluted in antibody diluent were applied and incubated at 4 °C overnight; the primary antibodies were as follows: PTGS2 (ABclonal, A3560, rabbit, 1:200), IGFBP2 (Proteintech, 66,644-1-IG, mouse, 1:200), WNT1 (ABclonal, A2475, rabbit, 1:200), LEP (bioss, bs-0409R, rabbit, 1:200), GFAP (Proteintech, 60,190-1-IG, mouse, 1:1000), β -catenin (huilanbio, ABB3523, rabbit, 1:200), and NF-κB p65 (Cell Signaling Technology, 8,242, rabbit, 1:200), with EDTA-based retrieval used for all targets.

Techniques: Binding Assay

Journal: Frontiers in Nutrition

Article Title: Astragaloside IV modulates oxidative stress and osteoimmune–Wnt signaling in ovariectomized rats: an integrated study of RNA sequencing, molecular docking, and experimental validation

doi: 10.3389/fnut.2026.1785452

Figure Lengend Snippet: Multiplex immunofluorescence imaging and semi-quantification of target proteins in femoral trabecular bone. Representative multiplex immunofluorescence images showing DAPI (nuclei), target protein staining, and merged overlays in the femoral trabecular region from Sham, OVX, and OVX-AS-IV-M treated rats. Panels show staining for (A) NF-κB p65, (B) Wnt1, (C) Gfap, (D) β-catenin, (E) Igfbp2, (F) Lep, and (G) Ptgs2. Scale bar, 50 μm. The corresponding semi-quantification of mean fluorescence intensity (MFI) for each marker is presented to the right of each image panel, normalized to the Sham group (set to 1). Data are presented as mean ± SD with individual data points overlaid. Statistical comparisons were performed using one-way ANOVA followed by Dunnett’s multiple-comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies diluted in antibody diluent were applied and incubated at 4 °C overnight; the primary antibodies were as follows: PTGS2 (ABclonal, A3560, rabbit, 1:200), IGFBP2 (Proteintech, 66,644-1-IG, mouse, 1:200), WNT1 (ABclonal, A2475, rabbit, 1:200), LEP (bioss, bs-0409R, rabbit, 1:200), GFAP (Proteintech, 60,190-1-IG, mouse, 1:1000), β -catenin (huilanbio, ABB3523, rabbit, 1:200), and NF-κB p65 (Cell Signaling Technology, 8,242, rabbit, 1:200), with EDTA-based retrieval used for all targets.

Techniques: Multiplex Assay, Immunofluorescence, Imaging, Staining, Fluorescence, Marker

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