anti mouse f4 80  (Cell Signaling Technology Inc)

Journal: Cell Death Discovery

Article Title: Excessive pyroptosis mediates the exacerbation of pneumonia caused by low-lethality influenza virus and secondary MRSA co-infection

doi: 10.1038/s41420-026-03031-z

Figure Lengend Snippet: A Flow cytometry analysis of the proportions of F4/80 + CD11b + recruitment macrophage in pulmonary after 6 h co-infection and B after 48 h co-infection. C Macrophages were deleted by clodronate liposomes which were administrated intranasally or intraperitoneally. D Flow cytometry analysis of Annexin V + PI + ‌late apoptosis and ‌necroptosis cells. ( n = 4) E Flow cytometry analysis of caspase-1 + PI + pyroptosis macrophage in different groups after 6 h co-infection and F after 48 h co-infection. Data are mean ± standard error of the mean (* p < 0.05; ** p < 0.01; *** p < 0.001) ( n = 4). IAV Influenza virus, MRSA methicillin-resistant S. aureus, CL clodronate liposomes.

Article Snippet: Then the samples were stained with fluorophore-conjugated anti-mouse antibodies or dyes [CoraLite® Plus 405 Anti-Mouse CD11c (Proteintech, Cat No. CL405-65602); anti-mouse CD170 (Siglec-F) Antibody (BioLegend, Cat#155507); anti-mouse CD45 Antibody (ABclonal, Cat A27305); FAM-YVAD-FMK Caspase-1 inhibitor (Immunochemistry, Lot#22N19); anti-mouse F4/80 Antibody (ABclonal, A27478); anti-mouse CD11b Antibody (BioLegend, Cat#101205); PI (BioLegend, Cat#421301); Annexin V-FITC (Beyotime, C1062M)] and fixed with 4% paraformaldehyde.

Techniques: Flow Cytometry, Infection, Liposomes, Virus

Journal: Cell Death Discovery

Article Title: Excessive pyroptosis mediates the exacerbation of pneumonia caused by low-lethality influenza virus and secondary MRSA co-infection

doi: 10.1038/s41420-026-03031-z

Figure Lengend Snippet: A RAW264.7 was infected with IAV and MRSA. Cell lysates were immunoblotted with anti-caspase-1 and anti-α-tubulin antibodies. Relative protein levels were calculated. ( n = 3) B Cell lysates were immunoblotted with anti-GSDMD and anti-GAPDH antibodies. ( n = 4) C Cell lysates were immunoblotted with anti-α-tubulin and anti-IL-1β antibodies. ( n = 4) D Immunofluorescence staining of F4/80 (red) and GSDMD (green), the scale bars are 20 μm(left) or 10 μm(right). Data are mean ± standard error of the mean (* p < 0.05; ** p < 0.01; *** p < 0.001) ( n = 3). IAV Influenza virus, MRSA methicillin-resistant S. aureus , GSDMD-FL Gasdermin D- full length, GSDMD-NT Gasdermin D-N terminal.

Article Snippet: Then the samples were stained with fluorophore-conjugated anti-mouse antibodies or dyes [CoraLite® Plus 405 Anti-Mouse CD11c (Proteintech, Cat No. CL405-65602); anti-mouse CD170 (Siglec-F) Antibody (BioLegend, Cat#155507); anti-mouse CD45 Antibody (ABclonal, Cat A27305); FAM-YVAD-FMK Caspase-1 inhibitor (Immunochemistry, Lot#22N19); anti-mouse F4/80 Antibody (ABclonal, A27478); anti-mouse CD11b Antibody (BioLegend, Cat#101205); PI (BioLegend, Cat#421301); Annexin V-FITC (Beyotime, C1062M)] and fixed with 4% paraformaldehyde.

Techniques: Infection, Immunofluorescence, Staining, Virus

Journal: Cell Death Discovery

Article Title: Excessive pyroptosis mediates the exacerbation of pneumonia caused by low-lethality influenza virus and secondary MRSA co-infection

doi: 10.1038/s41420-026-03031-z

Figure Lengend Snippet: A Representative pathological images (Scale bars: above 200 μm; below 100 μm) of lung tissue in different groups. Stain areas of images were calculated using image j ( n = 3). B Gross pathological changes of lung tissues were evaluated. C Inflammatory cytokines IL-1β, IL-6 and TNF-α mRNA levels were measured by qRT-PCR ( n = 5). Data are normalized to β-actin and presented as fold of gene expression in the Dsf groups compared to the coinfected groups. D Bacterial burden was determined by spreading 5 μL of the lung homogenates on the selective S. aureus medium ( n = 4). E Flow cytometry analysis of F4/80 + CD11c + tissue-resident macrophages. Data are mean ± standard error of the mean (* p < 0.05; ** p < 0.01; *** p < 0.001). IAV, Influenza virus; MRSA, methicillin-resistant S. aureus ; Dsf, disulfiram.

Article Snippet: Then the samples were stained with fluorophore-conjugated anti-mouse antibodies or dyes [CoraLite® Plus 405 Anti-Mouse CD11c (Proteintech, Cat No. CL405-65602); anti-mouse CD170 (Siglec-F) Antibody (BioLegend, Cat#155507); anti-mouse CD45 Antibody (ABclonal, Cat A27305); FAM-YVAD-FMK Caspase-1 inhibitor (Immunochemistry, Lot#22N19); anti-mouse F4/80 Antibody (ABclonal, A27478); anti-mouse CD11b Antibody (BioLegend, Cat#101205); PI (BioLegend, Cat#421301); Annexin V-FITC (Beyotime, C1062M)] and fixed with 4% paraformaldehyde.

Techniques: Staining, Quantitative RT-PCR, Gene Expression, Flow Cytometry, Virus

Journal: Cell Death Discovery

Article Title: Excessive pyroptosis mediates the exacerbation of pneumonia caused by low-lethality influenza virus and secondary MRSA co-infection

doi: 10.1038/s41420-026-03031-z

Figure Lengend Snippet: A Experimental scheme for assessing the combination treatment of oseltamivir, linezolid and disulfiram. B Gross pathological changes of lung tissues were evaluated. C Lung index was calculated in different groups ( n = 6). Lung index = lung weight of mice (mg)/body weight of mice (g). D mRNA levels of IL-1β, CXCL-2 and TNF-α were measured by qRT-PCR ( n = 4). Data are normalized to β-actin and presented as fold of gene expression in the infected groups compared to the control. E Representative pathological images (Scale bars: above 200 μm; below 100 μm) of lung tissue in different groups. Stain areas of images were calculated using image j and then normalized to the average area of the control group ( n = 3). F Immunofluorescence staining of F4/80 (red) and GSDMD (green), the scale bars are 20 μm (left) or 10 μm (right). Data are mean ± standard error of the mean (* p < 0.05; ** p < 0.01; *** p < 0.001). IAV Influenza virus, MRSA methicillin-resistant S. aureus, Ost oseltamivir, Lzd linezolid, Dsf disulfiram.

Article Snippet: Then the samples were stained with fluorophore-conjugated anti-mouse antibodies or dyes [CoraLite® Plus 405 Anti-Mouse CD11c (Proteintech, Cat No. CL405-65602); anti-mouse CD170 (Siglec-F) Antibody (BioLegend, Cat#155507); anti-mouse CD45 Antibody (ABclonal, Cat A27305); FAM-YVAD-FMK Caspase-1 inhibitor (Immunochemistry, Lot#22N19); anti-mouse F4/80 Antibody (ABclonal, A27478); anti-mouse CD11b Antibody (BioLegend, Cat#101205); PI (BioLegend, Cat#421301); Annexin V-FITC (Beyotime, C1062M)] and fixed with 4% paraformaldehyde.

Techniques: Quantitative RT-PCR, Gene Expression, Infection, Control, Staining, Immunofluorescence, Virus

Journal: Advanced Science

Article Title: FAPα + Macrophages Orchestrate Immune Evasion in Multiple Myeloma by Dual Regulation of PD‐L1 and T Cell Senescence

doi: 10.1002/advs.202506239

Figure Lengend Snippet: FAPα Inhibition Overcomes Immunotherapy Resistance in MM by Depleting TAMs and Enhancing GZMB‐Mediated Cytotoxicity. (A–C) Representative images of 5TGM1 tumors (A), tumor growth curves (B), and tumor weight (C) from the indicated treatment groups. (D‐E) IHC staining (D) and corresponding IOD analysis (E) of CD138, F4/80, FAPα, PD‐L1, CD4, CD8, IFNγ, and GZMB in 5TGM1 tumors (Magnification ×200. Scale bar, 100 µm). Data are presented as mean ± SD. Each dot means independent samples. ns, no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Statistical analysis was performed using a 2‐way ANOVA test in B, a 2‐tailed Student's t ‐test in C and E. FAPα, fibroblast activation protein alpha; PD‐L1, programmed cell death‐ligand 1; PD‐1, programmed cell death protein‐1; IHC, Immunohistochemistry; IOD, Integrated Optical Density; IFNγ, Interferon gamma; GZMB, granzyme B.

Article Snippet: The used antibodies were: anti‐human CD138 (Cat # BX500300 , Biolynx, China), anti‐human CD68 (Cat #BX50031, AB_2936308, Biolynx, China), anti‐human FAP (Cat #BMS168, AB_10597443, eBioscience, USA), anti‐human/mouse CD138 (Cat #ab128936, AB_11150990, Abcam, USA), anti‐mouse F4/80 (Cat #70076, AB_2799771, Cell Signaling Technology, USA), anti‐mouse FAP (Cat #PA5‐99313, AB_2818246, Invitrogen, USA), anti‐mouse PD‐L1 (Cat #17952‐1‐AP, AB_10597552, Proteintech, USA), anti‐mouse CD4 (Cat #ab183685, AB_2686917, Abcam, USA), anti‐mouse CD8 (Cat #ab209775, AB_2860566, Abcam, USA), anti‐mouse IFNγ (Cat #105995‐T08, Sino Biological inc. China) and anti‐mouse GZMB (Cat #48298, Signalway Antibody, USA).

Techniques: Inhibition, Immunohistochemistry, Activation Assay

Journal: Advanced Science

Article Title: FAPα + Macrophages Orchestrate Immune Evasion in Multiple Myeloma by Dual Regulation of PD‐L1 and T Cell Senescence

doi: 10.1002/advs.202506239

Figure Lengend Snippet: FAPα Inhibition Potentiates Anti‐PD‐1/PD‐L1 Therapy Across Multiple Tumor Types. (A) Correlation analysis of FAPα mRNA expression and PFI / OS in pan‐cancer data. (B) Correlation analysis of FAPα expression and immune cell infiltration in the tumor microenvironment. (C–E) Representative images(C), tumor growth curves(D), and tumor weight (F) of EL4 tumors from the indicated groups treated by αPD‐1/αPD‐L1 antibodies combined with or without FAPα inhibitor. (F,G) Representative images (F) and IOD analysis of IHC staining (G) of Ki67, F4/80, FAPα, PD‐L1, CD4, CD8, IFNγ, and GZMB in EL4 tumors (Magnification ×200. Scale bar, 100 µm). (H–J) Representative images(H), tumor growth curves(I), and tumor weight (J) of CT26 tumors from the indicated groups treated by αPD‐1/αPD‐L1 antibodies combined with or without FAPα inhibitor. (K, L) Representative images (K) and IOD analysis of IHC staining (L) of Ki67, F4/80, FAPα, PD‐L1, CD4, CD8, IFNγ, and GZMB in CT26 tumors (Magnification ×200. Scale bar, 100 µm). Data are presented as mean ± SD. Each dot means independent samples. ns, no significant difference. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Statistical analysis was performed using log‐rank test in A, a 2‐way ANOVA test in D and I,2‐tailed Student's t ‐test in E, G, J, and L. FAPα, fibroblast activation protein alpha; PFI, progression‐free interval; OS, overall survival; IHC, Immunohistochemistry; IOD, Integrated Optical Density; PD‐L1, programmed cell death‐ligand 1; PD‐1, programmed cell death protein‐1; IFNγ, Interferon gamma; GZMB, granzyme B.

Article Snippet: The used antibodies were: anti‐human CD138 (Cat # BX500300 , Biolynx, China), anti‐human CD68 (Cat #BX50031, AB_2936308, Biolynx, China), anti‐human FAP (Cat #BMS168, AB_10597443, eBioscience, USA), anti‐human/mouse CD138 (Cat #ab128936, AB_11150990, Abcam, USA), anti‐mouse F4/80 (Cat #70076, AB_2799771, Cell Signaling Technology, USA), anti‐mouse FAP (Cat #PA5‐99313, AB_2818246, Invitrogen, USA), anti‐mouse PD‐L1 (Cat #17952‐1‐AP, AB_10597552, Proteintech, USA), anti‐mouse CD4 (Cat #ab183685, AB_2686917, Abcam, USA), anti‐mouse CD8 (Cat #ab209775, AB_2860566, Abcam, USA), anti‐mouse IFNγ (Cat #105995‐T08, Sino Biological inc. China) and anti‐mouse GZMB (Cat #48298, Signalway Antibody, USA).

Techniques: Inhibition, Expressing, Immunohistochemistry, Activation Assay

Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: TREM2 + TAMs hinder the therapeutic effectiveness of GPC3-CAR-T cells (A) UMAP visualization of scRNA-seq data from tumor biopsies following IL15-GPC3-CAR-T therapy (data from Steffin et al., Nature 2025; GEO: GSE253352 ). (B) The heatmap displayed the signature differentially expressed genes across distinct myeloid cell subpopulations. (C) The bar chart showed the proportions of myeloid cell subsets in the PD and PR groups. (D) The boxplot demonstrated the statistical differences among various myeloid subpopulations. (E) In vitro cytotoxicity measured by lactate dehydrogenase (LDH) release at indicated effector cell vs. target cell ratios (E:T ratios) using Hepa1-6 cells overexpressing human GPC3 or vector control were co-cultured with Ctrl-CAR-T (targeted CD19) or GPC3-CAR-T cells. (F) Subcutaneous tumor growth and survival in C57BL/6J mice ( n = 6/group) inoculated with GPC3-Hepa1-6 cells and treated with Ctrl- or GPC3-CAR-T cells (1 × 10 6 ) via tail vein. (G) Orthotopic liver tumor model: tumor weight at day 28 and survival in mice receiving intrahepatic GPC3-Hepa1-6 implants followed by CAR-T treatment. (H) Flow cytometric analysis of immune and stromal cell populations in tumors from subcutaneous and orthotopic models following GPC3-CAR-T treatment. (I) Expression of TAM-associated genes (Trem2, Spp1, Cd274, Arg1, Tgfb1, and Cd206) in F4/80 + TAMs sorted from tumors of GPC3-CAR-T-treated mice, measured by RT-qPCR. (J) Schematic of the orthotopic HCC model for CAR-T therapy in Trem2 -WT/KO mice. (K) Tumor representative photographs of different groups were shown. Dot plots showed liver weight, liver-to-body weight ratio, per liver, n = 8. (L) UMAP plots of individual groups displayed distinct intratumoral cell populations. n = 2 for all treatment groups on scRNA sequence. (M) Proportion of the distinct intratumoral cell populations. Data were represented by mean ± SEM. ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rabbit anti-mouse F4/80 antibody , CST , Cat#70076.

Techniques: In Vitro, Plasmid Preparation, Control, Cell Culture, Expressing, Quantitative RT-PCR, Sequencing

Journal: Nature Communications

Article Title: Living sensor display implanted on skin for long-term biomarker monitoring

doi: 10.1038/s41467-025-67384-2

Figure Lengend Snippet: a Bright-field images of the engrafted tissue-engineered skin before LPS stimulation, more than 2 months post-transplantation. b Fluorescence images of the engrafted tissue-engineered skin before LPS administration, c 2 days after the intraperitoneal administration of LPS, d 7 days after administration, e Fluorescence image of the engrafted tissue-engineered skin at 2 days after subcutaneous administration of LPS on the 9th day post-initial administration. f Time course of fluorescence intensity calculated from the image analysis of the tissue-engineered skin. The vertical dotted lines indicate the time points of fluorescence observation, and the downward‑pointing triangles indicate the timing of LPS administration. g Comparison of fluorescence intensities at 7 days and 9 days after administration. Data are presented as mean ± SD from n = 5 biologically independent mice. p = 0.00000056. The statistical analysis was performed using t-tests (two-tailed). *** p < 0.001. h Comparison of fluorescence intensities at 2 days and 7 days after administration. The statistical analysis was performed using t-tests (two-tailed). Data are presented as mean ± SD from n = 5 biologically independent mice. p = 0.3898 i Immunostaining image merged for human mitochondria and GFP from a skin section taken from around the engrafted skin at 9 days after LPS administration. Representative of three independent experiments with similar results. j Fluorescence image confirming the localisation of inflammatory cells positive for Ly6g and F4/80 in the magnified area. Representative of three independent experiments with similar results.

Article Snippet: Following the removal of the BSA solution, the tissue sections were incubated with the diluted primary antibody: Cytokeratin 14 Polyclonal antibody (Proteintech・10143-1-AP,CK14), Anti-Collagen IV antibody(abcam・ab6586, Col4),Vimentin (D21H3) XP Rabbit mAb (CST 5741S, Vimentin), Anti-GFP (Rat IgG2a) Monoclonal (GF090R) CC (nacalali,04404-84,GFP), Anti-GFP Antibody (abcam・ab290, GFP), Purified Rat Anti-Mouse CD31 (BD pharmingen・553370, CD31), Anti-Mitochondria antibody (Merck (Sigma)・MAB1273, Human Mitochondria), Rat Anti-Mouse F4/80 antibody (Bio-Rad・MCA497GA, F4/80), Anti-CD86 antibody (Cell Signalling Technology・ #19589S, CD86) Anti-CD163 antibody (abcam・ab182422, CD163), Anti-Ly6g antibody (abcam・ab25377, Ly6g), overnight at 4 °C within the moisture chamber.

Techniques: Transplantation Assay, Fluorescence, Comparison, Two Tailed Test, Immunostaining