horse radish peroxidase conjugated immpress anti mouse igg  (Vector Laboratories)

Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Article Snippet: The anti-mCII ELISA kits were used according to manufacturer's protocol (2036T; Chondrex).

Techniques: Indirect ELISA

Journal: Bioactive Materials

Article Title: Glucose/ROS-responsive and redox-gated adaptive hydrogel dressing for accelerating diabetic wound repair via synergistic cGAS/STING pathway inhibition and oxidative stress alleviation

doi: 10.1016/j.bioactmat.2026.03.025

Figure Lengend Snippet: Angiogenesis and collagen deposition in diabetic wound tissues following HPSL@SG hydrogel treatment. (A) Dihydroethidium (DHE) immunofluorescence staining and (B) semi-quantitative analysis of wound tissues from each treatment group on day 7, scale bar = 100 μm. Immunofluorescence staining of (C) MMP-9, IL-6, and IL-10, and (D) CD31, VEGF-A, and collagen I in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. (E-J) Mean relative fluorescence intensity of each indicator in wound tissue sections from each treatment group on day 7, scale bar = 100 μm. All data are shown as mean ± SEM (n = 6).

Article Snippet: IL-6 and IL-10-specific antibodies were purchased from Bosterbio (Wuhan, China).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Integrative Medicine Research

Article Title: Therapeutic effects of pomegranate hot-water extract via inhibition of apoptosis and oxidative stress in a DHEA-induced mouse model of PCOS

doi: 10.1016/j.imr.2025.101264

Figure Lengend Snippet: Pg-hWE alleviates PCOS symptoms in a mouse model based on H&E staining. (A) PCOS mouse model construction. The mice were subcutaneously administered with 30 mg/kg of DHEA for six weeks. One week prior to start of SC administration, each group was administered the appropriate drug (tap water, dextrin, and Pg-hWE) orally. (B) Body weight was measured once a week. (C) Results of the AMH ELISA with mouse serum. (D) Mouse ovary histology by H&E staining. (E) Results of ovarian follicles by type. # p < 0.05 ## p < 0.01 compared with the Normal group, * p < 0.05 and ⁎⁎ p < 0.01 compared with the PCOS group.

Article Snippet: ELISA of mouse serum was performed using the CUSABIO's anti-Müllerian hormone ELISA kit (CUSABIO, Houston, TX, USA) as reported previously, and all preparation and experimental procedures were performed according to the protocol provided by CUSABIO.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Gut Microbes

Article Title: Phenotypic profiling of neutrophils in acute Clostridioides difficile infection identifies a TNF-induced activation signature associated with epithelial damage

doi: 10.1080/19490976.2026.2659430

Figure Lengend Snippet: Single-cell transcriptomics reveal an increase in a hyperactivated neutrophil cluster in response to CDI. (A) Experimental plan: Colon tissue was collected from a mouse challenged with C. difficile (1 × 10 6 M7404 spores) during the acute phase of infection (i.e., 2 d after challenge). Neutrophils were enriched using MACS negative selection and submitted for 10× genomics. (B) Uniform Manifold Approximation and Projection (UMAP) of colon neutrophil clusters. (C) Percent distribution of colonic neutrophil clusters. (D) Heatmap of differentially expressed genes for each cluster. (E) Differentially expressed genes from each colon neutrophil cluster of the single-cell dataset were used as the input for Metascape pathway enrichment analysis. (F and G) Module score of colon neutrophils for (F) genes associated with phagocytosis, neutrophil activation, degranulation, and ROS production, and G) genes associated with human CDI and colitis. For (F), gene lists were obtained from prior literature and Gene Ontology database ( Xie et al. 2020 and the Gene Ontology Browser on Mouse Genome Informatics website: https://www.informatics.jax.org/mgihome/GO/project.shtml ). For (G), human gene-disease association lists were obtained from MalaCards: Clostridium difficile colitis (DOID:0060185) and Colitis (DOID:0060180). For box plots, the middle line represents median values of gene expression, the upper and lower hinges of box represent first and third quartile (25th and 75th percentiles), and the upper/lower whiskers extend from the hinge to the largest/smallest value no further than 1.5*interquartile range (IQR) from the hinge (where IQR is the distance between the first and third quartiles). Stats: two-tailed Wilcoxon rank-sum test (F and G); * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: Neutrophils were enriched using MACS MS columns (Miltenyi #130-097-658) and stained with antibodies for Ly6G (BioLegend #127654 or #563978), CD11b (BioLegend #101259 or Thermo Fisher #12-0112-82), and viability dye (Thermo Fisher # L10120 ).

Techniques: Single-cell Transcriptomics, Infection, Selection, Single Cell, Activation Assay, Gene Expression, Two Tailed Test

Journal: Gut Microbes

Article Title: Phenotypic profiling of neutrophils in acute Clostridioides difficile infection identifies a TNF-induced activation signature associated with epithelial damage

doi: 10.1080/19490976.2026.2659430

Figure Lengend Snippet: CDI drives systemic neutrophil transcriptomic alterations that contribute to tissue injury. (A) Experimental plan: Bone marrow and blood samples were collected from an uninfected mouse and a mouse challenged with C. difficile (1 × 10 6 M7404 spores) on day 2 after infection. Neutrophils were enriched using MACS negative selection and submitted for 10× genomics. (B) UMAP of bone marrow and blood neutrophil clusters. (C) Percent distribution of bone marrow and blood neutrophil clusters. (D) Waterfall plot of gene set enrichment analysis of bone marrow and blood neutrophils: comparison between data from uninfected and infected mouse. (E and F) Module scoring of bone marrow and blood neutrophils from uninfected and infected mouse for genes associated with (E) granule protein, NADPH oxidase formation, and ROS production; and (F) with human CDI. For (E), gene lists were used from the Reactome database, and for (F), gene lists were curated from MalaCards and from Tsakiroglou et al. 2024 . For box plots, the middle line represents median values of gene expression, the upper and lower hinges of box represent first and third quartile (25th and 75th percentiles), and the upper/lower whiskers extend from the hinge to the largest/smallest value no further than 1.5*interquartile range (IQR) from the hinge (where IQR is the distance between the first and third quartiles). Stats: two-tailed Wilcoxon rank-sum test (E and F); * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: Neutrophils were enriched using MACS MS columns (Miltenyi #130-097-658) and stained with antibodies for Ly6G (BioLegend #127654 or #563978), CD11b (BioLegend #101259 or Thermo Fisher #12-0112-82), and viability dye (Thermo Fisher # L10120 ).

Techniques: Infection, Selection, Comparison, Gene Expression, Two Tailed Test

Journal: Gut Microbes

Article Title: Phenotypic profiling of neutrophils in acute Clostridioides difficile infection identifies a TNF-induced activation signature associated with epithelial damage

doi: 10.1080/19490976.2026.2659430

Figure Lengend Snippet: cNeu3-like neutrophils share a transcriptomic profile with TNF-polarized neutrophils during acute CDI, and TNF treatment induces this phenotype in vitro . (A) Monocle pseudotime trajectory analysis of bone marrow and blood neutrophil clusters from uninfected and infected mouse. (B) Cell fate analysis performed using Monocle3 by analyzing genes upregulated as cells transition from Neu5 to Neu6 along the pseudotime trajectory. Each point represents a cell in UMAP space, and cells are colored by the gene module upregulated as cells transition from Neu5 to Neu6. (C) Metascape KEGG pathway enrichment analysis of genes upregulated as cells transition from Neu5 to Neu6. (D) Neutrophil cytokine-induced polarization cellular state plots of bone marrow, blood, and colon neutrophils from uninfected and C. difficile -infected mouse. DEGs from neutrophils polarized with TNF, type I/II interferons, IL-1α/β, or GM-CSF were utilized to define neutrophil transcriptomic states induced by the corresponding cytokines. For this analysis, gene lists were obtained from Cui et al. 2023. (E) Cui 2023 TNF-polarized neutrophil gene signature and KEGG TNF signaling pathway signature for Neu6, Neu8, and cNeu3 plotted against pseudotime. (F) Amount of TNF (pg/mg of protein) collected from uninfected and C. difficile -infected mice during acute CDI (i.e., day 2 after challenge). (G) Experimental plan: Neutrophils were isolated from the bone marrow of WT mice and enriched by MACS bead-based negative selection. Neutrophils were plated at 1 × 10 6 cells/well in 96-well plates and then treated with recombinant murine TNF (50 ng/ml) or control (PBS) for 2 h. Cells were incubated at 37 °C and then stained for flow cytometry. (H) Representative flow cytometry plot showing CD11b and IL-1β staining in untreated and TNF-treated samples. (I) Percent distribution of CD11b hi IL-1β hi in untreated and TNF-treated samples. (J) Comparison of MALT1, MIP1α, and CD63 gMFI in gate 1 vs. gate 2 populations in untreated and TNF-treated samples. Panels (F) and (J and K) are presented as box-and-whisker plots (boxes = interquartile range, whiskers = minimum and maximum, line = median). Panel (F) shows data combined from 4 independent experiments, n = 14 (control) and 18 (CDI) (F); panels (H) and (J) are representative of 2 independent experiments, n = 3 per group; panel I is combined data points from two independent experiments, total n = 6 per group. Stats: Mann–Whitney test (F); Spearman correlation (G); unpaired t -test with Welch's correction (I and J); * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: Neutrophils were enriched using MACS MS columns (Miltenyi #130-097-658) and stained with antibodies for Ly6G (BioLegend #127654 or #563978), CD11b (BioLegend #101259 or Thermo Fisher #12-0112-82), and viability dye (Thermo Fisher # L10120 ).

Techniques: In Vitro, Infection, Isolation, Selection, Recombinant, Control, Incubation, Staining, Flow Cytometry, Comparison, Whisker Assay, MANN-WHITNEY