vectashield antifade mounting media  (Vector Laboratories)

Journal: Cell

Article Title: Spatial biology reveals altered macrophage states in immunosuppressed non-melanoma skin cancer

doi: 10.1016/j.cell.2026.07.031

Figure Lengend Snippet: (A) Multiplex immunofluorescent analysis of cSCC tissue stained with antibodies against cytokeratin, CD4, CD8, CD68, FOXP3,LAG3 and PD-L1 and counterstained with DAPI. CK, cytokeratin. (B) Violin plots demonstrating the density distributions of immune cell phenotypes (cells/mm 2 , log 2 ) in tumor and stromal regions of cSCC samples from immunosuppressed and immunocompetent patients. (C) Heat map clinical and demographics annotation plot of patients in the external validation cohort from Cleveland Clinic. IS, immunosuppression; HM, hematological malignancy; OTR, organ transplant recipient; HN, head and neck; cSCC, cutaneous squamous cell carcinoma; (D) Multiplex immunofluorescent analysis of cSCC tissue from IC and IS patients in the Cleveland Clinic cohort stained with antibodies against CD11b, CD11c, CD40, CD68, and HLA-DR, together with cytokeratin (Pan-CK) and DAPI counterstain. (E) Violin plots showing the density distributions (cells/mm 2 , log 2 ) of total myeloid cells (CD11b + CD11c + CD68 + ), macrophages (CD68 + ), dendritic cells (DCs; CD11c + CD68 − ), and antigen-presenting DCs (CD11c + CD68 − HLA-DR + ) in IC and IS tumors as quantified by multiplex immunofluorescence. Mean ± SEM is indicated; p values were determined by two-tailed Student’s t-test. (F) UMAP visualization of macrophage populations from all IC & IS NMSC using scRNA-seq, annotated by transcriptionally defined TAM subtypes: resident-tissue macrophages (RTMs), proliferative TAMs (Prof-TAMs), IFN-responsive TAMs (IFN-TAMs), lipid-associated TAMs (LA-TAMs), ECM-degradation TAMs (ECM-Deg), and inflammatory cytokine-enriched TAMs (Inflam-TAMs).(G) Dot plot of the canonical gene markers used to identify the macrophages cell types (H) Bar graph comparing the proportions of TAM populations from IC & IS NMSCs (I) Dot plot showing the difference in the macrophage subtypes between IS and IC with corresponding FDR values. (J) Number of differentially expressed genes (DEGs) between IS & IC tumors across the macrophages subclusters. (K) Pathway enrichment analysis of DEGs between IS and IC TAM populations showing key immunoregulatory and inflammatory signaling pathways; circle size represents significance (p value) and color indicates z-score of pathway activation. Pos, positive; reg, regulation; Ag, antigen.

Article Snippet: Counterstaining of the cell nuclei and coverslipping was performed using VectaShield Antifade Mounting Media with DAPI (Vector Laboratories, Newark, CA).

Techniques: Profiling, Multiplex Assay, Analysis, Tissue, Staining, Immune Cell, Biomarker Discovery, Immunofluorescence, Two Tailed Test, Protein-Protein interactions, Activation Assay

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: Citrullinated and Homocitrullinated Proteins/Peptides are Present in DBA/1J Metatarsophalangeal Joints. A) Representative hematoxylin + eosin (H&E) at 80× magnification and immunofluorescent micrographs at 20× magnification of joints from DBA/1J mice immunized with bovine type II collagen (arthritic: N = 9; non-arthritic: N = 10) or PBS (N = 9). Scale bars represent 500 μm. Max projections of Z-stacks show nuclei (DAPI; blue), citrullinated proteins/peptides (CitP; green), and homocitrullinated proteins/peptides (HomoCitP; red). Corrected total fluorescence intensity for B) CitP and C) HomoCitP in three joint structures: bone marrow, synovium, and cartilage. Graphs show the median [IQR], with each symbol representing an individual mouse. Statistical analysis was performed using Kruskal-Wallis with Dunn's multiple comparisons test, and the resulting p-values were p = 0.0046, p = 0.0084, p = 0.4076, p = 0.0143, p = 0.0136, and p = 0.1733 respectively, with p-values for pairwise comparisons indicated on the graphs. ns: not significant.

Article Snippet: Following application of TrueView autofluorescence quencher, slides were mounted using Vectashield Vibrance Antifade mounting media with DAPI (SP-8500-15; Vector Laboratories; USA) and air dried for 2 h before imaging on the Nikon Ti2-E microscope.

Techniques: Fluorescence