Journal: Translational Cancer Research

Article Title: Phagocytic remodeling in CD74 High tumor-associated macrophages during brain metastasis of lung adenocarcinoma

doi: 10.21037/tcr-2026-1-0228

Figure Lengend Snippet: Identification and characterization of CD74 High TAM subpopulations in the LT and BM microenvironments. (A) Circular UMAP visualization displaying the cluster distribution, with outer sectors quantifying cell counts and proportions for each subset. (B,C) Density plots highlighting the expression patterns of the macrophage lineage marker CD68 (B) and CD74 (C), where green intensity indicates high expression levels. (D) Validation of CD74 and CD68 co-localization in BM tissue sections via multiplex immunofluorescence staining. Representative images show the merged and single-channel expression (CD74, CD68, DAPI) at 20× magnification (scale bar: 50 µm, left panels), alongside a high-resolution view of the indicated region at 50× magnification (scale bar: 20 µm, right panels). (E) UMAP visualization of TAMs stratified by tissue origin and CD74 expression levels, categorized into five distinct subsets: LT_High (20.1%), LT_Low (7.5%), BM_High (5%), BM_Low (17.7%), and intermediate (50%). (F) Volcano plot illustrating the differential gene expression analysis between the LT_High and BM_High subsets. Red dots denote genes significantly upregulated in LT_High TAMs, while blue dots represent downregulated genes (top 10 DEGs are labeled). BM, brain metastasis; DAPI, 4',6-diamidino-2-phenylindole; DEG, differentially expressed genes; LT, lung tumor; TAM, tumor-associated macrophage; UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: Nuclear counterstaining and mounting were conducted using 4',6-diamidino-2-phenylindole (DAPI) Fluoromount-GTM (Cat. No. 36308ES20; Yeasen, Shanghai, China).

Techniques: Expressing, Marker, Biomarker Discovery, Multiplex Assay, Immunofluorescence, Staining, Gene Expression, Labeling

Journal: Virus Research

Article Title: Characterization and antibody preparation of the gene products of grouper iridovirus ORF120L

doi: 10.1016/j.virusres.2025.199625

Figure Lengend Snippet: Immunofluorescence staining analysis of GIV-120L. GK cells were infected with GIV and the localization of GIV-120L expression at 0, 12, and 24 hpi was analyzed by immunofluorescence staining using monoclonal antibodies targeting GIV-120L-His. Nuclei were counterstained with DAPI. Fluorescence signals were visualized using a fluorescence microscope (Zeiss, Germany). Bright-field images show cell morphology; FITC indicates location of target protein; DAPI indicates location of DNA; white arrow indicates presumed location of viral DNA.

Article Snippet: The wells were washed with PBS, and blocking was performed by adding 500 μL of 10 % (v/v) FBS/PBS for 1 h. The wells were washed with PBS, and 500 μL of primary antibody (mouse serum in 10 % (v/v) FBS/PBS at a dilution of 1:100) was added and the mixture incubated at 37 °C for 1 h. The wells were then washed with PBS, and 500 μL of secondary antibody (goat anti-mouse IgG-FITC in 10 % (v/v) FBS/PBS at a dilution of 1:300) was added and incubated at 37 °C for 1 h. The wells were washed with PBS, and the coverslips were recovered, mounted with DAPI Fluoromount-GTM (Southern Biotech, USA), and observed and imaged using a fluorescence microscope (Zeiss, Germany).

Techniques: Immunofluorescence, Staining, Infection, Expressing, Bioprocessing, Fluorescence, Microscopy

Journal: Frontiers in Immunology

Article Title: IFIT3 activation significantly contributes to HIV-1-associated neurodegenerative disorder-mediated neuroinflammation

doi: 10.3389/fimmu.2025.1532318

Figure Lengend Snippet: Immunocytochemical analysis of IFIT3 and STAT1 protein expression in differentiated SH-SY5Y cells. Immunocytochemical analysis of IFIT3 and STAT1 expression in differentiated SH-SY5Y cells following HIV Tat and cART treatments. (A, D) Representative confocal microscopy images showing IFIT3 and STAT1 protein expression in cells treated with HIV Tat protein, combination cART, or Tat+cART for 24 hours. IFIT3 and STAT1 were detected using Alexa Fluor 488-conjugated antibodies (green), and nuclei were counterstained with DAPI (blue). Phase contrast images detail cellular morphology, and composite images combine all fluorescence channels. (B, E) Quantitative analysis of IFIT3 and STAT1 expression under different treatment conditions. Data were collected from three independent experiments, with 5–6 images per experiment and 5–10 cells analyzed per image. (C, F) Machine learning analysis evaluating classification accuracy based on IFIT3 and STAT1 expression patterns. Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparisons test. *P < 0.05; **P < 0.0027; ****P < 0.0001; ns, not significant.

Article Snippet: Nuclear staining and mounting of the cell nuclei were performed with Fluoromount-GTM DAPI (catalog# 010001, Southern Biotech), and the coverslips were mounted on microscope slides.

Techniques: Expressing, Confocal Microscopy, Fluorescence

Journal: Frontiers in Immunology

Article Title: IFIT3 activation significantly contributes to HIV-1-associated neurodegenerative disorder-mediated neuroinflammation

doi: 10.3389/fimmu.2025.1532318

Figure Lengend Snippet: (A) Immunohistochemical analysis of cerebellum brain tissue from humanized mice subjected to HIV-1 and cART is shown in the remaining panels. Representative images illustrate IFIT3 expression (green fluorescence), neuronal cells labeled with the NeuN marker (red fluorescence), and nuclear staining with DAPI (blue fluorescence). Column 1 shows IFIT3 expression. Column 2, NeuN labeling; Column 3, DAPI staining; Column 4, a composite image of IFIT3, NeuN, and DAPI. Column 5 shows phase contrast (PC) imaging to detail the cellular morphology. Column 6 shows a composite overlay of IFIT3/NeuN/DAPI with phase contrast imaging to provide a holistic view of expression patterns within the cellular architecture. (B) shows the fluorescence intensity, and (C) shows the accuracy analysis via MATLAB. The study included brain tissue samples from six individual mice (n=6) in each treatment group.Differences in IFIT3 expression were statistically analyzed via one-way ANOVA with Dunnett’s multiple comparisons test. Significance levels are indicated as *p = 0.219, ****p < 0.0001 and ns, not significant.

Article Snippet: Nuclear staining and mounting of the cell nuclei were performed with Fluoromount-GTM DAPI (catalog# 010001, Southern Biotech), and the coverslips were mounted on microscope slides.

Techniques: Immunohistochemical staining, Expressing, Fluorescence, Labeling, Marker, Staining, Imaging