markers arg 1  (R&D Systems)

Journal: Cell Division

Article Title: FGF4 drives tumor progression in triple-negative breast cancer via IL6/STAT3-mediated macrophage M2 polarization and immune suppression

doi: 10.1186/s13008-025-00164-y

Figure Lengend Snippet: Immune cell infiltration characteristics and core gene correlation analysis in TNBC samples. A Bar chart of immune cell infiltration levels in TNBC and normal samples analyzed using the CIBERSORT algorithm for 22 immune cell types; B Comparison of immune cell infiltration levels between TNBC and normal samples using the CIBERSORT algorithm; C Spearman correlation analysis of the four core genes (CDH2, FGF4, HIST1H3C, SOX2) and their correlation with the infiltration levels of 22 immune cell types; D Spearman correlation analysis of FGF4 expression and its association with the expression of M2 macrophage-related genes (TGFB1, STAT3, CCL2, SMAD3, IL6, and NFKB1)

Article Snippet: After blocking with a blocking solution (Beyotime, China) for 1 h at room temperature, the sections were incubated overnight at 4 °C with primary antibodies against M2 macrophage markers Arg1 (1:200, Cell Signaling Technology, USA, 93668) and CD206 (1:200, BioLegend, USA, C068C2).

Techniques: Comparison, Expressing

Journal: Cell Division

Article Title: FGF4 drives tumor progression in triple-negative breast cancer via IL6/STAT3-mediated macrophage M2 polarization and immune suppression

doi: 10.1186/s13008-025-00164-y

Figure Lengend Snippet: Impact of FGF4 on the IL6/STAT3 axis and macrophage polarization. A The experimental workflow diagram illustrates the construction of FGF4 overexpressing and knockdown TNBC cell lines, co-culture with macrophages, and subsequent analysis procedures. B qRT-PCR analysis of FGF4 mRNA expression levels in MDA-MB-231 cells with FGF4 overexpression or knockdown. C Western blot analysis of STAT3, p-STAT3, and IL-6 expression in macrophages co-cultured with MDA-MB-231 cells under different FGF4 expression conditions. D Quantification of protein expression levels. E Immunofluorescence detection of M1 macrophage polarization markers (iNOS, CD86) in macrophages. F Quantification of iNOS fluorescence in each group. G Quantification of CD86 fluorescence in each group. H Immunofluorescence staining for M2 macrophage polarization markers (Arg1, CD206) in macrophages. I Quantification of CD206 fluorescence in each group. J Quantification of Arg1 fluorescence in each group. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were performed in triplicate

Article Snippet: After blocking with a blocking solution (Beyotime, China) for 1 h at room temperature, the sections were incubated overnight at 4 °C with primary antibodies against M2 macrophage markers Arg1 (1:200, Cell Signaling Technology, USA, 93668) and CD206 (1:200, BioLegend, USA, C068C2).

Techniques: Knockdown, Co-Culture Assay, Quantitative RT-PCR, Expressing, Over Expression, Western Blot, Cell Culture, Immunofluorescence, Fluorescence, Staining

Journal: Cell Division

Article Title: FGF4 drives tumor progression in triple-negative breast cancer via IL6/STAT3-mediated macrophage M2 polarization and immune suppression

doi: 10.1186/s13008-025-00164-y

Figure Lengend Snippet: Role of the IL6/STAT3 signaling pathway in FGF4-mediated macrophage polarization. A – C Flow cytometry analysis of M2 polarization markers (Arg1, CD206) expression and quantitative analysis in macrophages. D – F Flow cytometry analysis of M1 polarization markers (CD68, iNOS) expression and quantitative analysis in macrophages. G – I Immunofluorescence analysis of M2 polarization markers (iNOS, CD86) expression and quantitative analysis in macrophages. (J-L) Immunofluorescence analysis of M1 polarization markers (CD68, iNOS) expression and quantitative analysis in macrophages. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were performed in triplicate

Article Snippet: After blocking with a blocking solution (Beyotime, China) for 1 h at room temperature, the sections were incubated overnight at 4 °C with primary antibodies against M2 macrophage markers Arg1 (1:200, Cell Signaling Technology, USA, 93668) and CD206 (1:200, BioLegend, USA, C068C2).

Techniques: Flow Cytometry, Expressing, Immunofluorescence

Journal: Cell Division

Article Title: FGF4 drives tumor progression in triple-negative breast cancer via IL6/STAT3-mediated macrophage M2 polarization and immune suppression

doi: 10.1186/s13008-025-00164-y

Figure Lengend Snippet: Immune suppressive function of FGF4-regulated macrophage M2 polarization and its impact on MDA-MB-231 breast cancer cell proliferation, migration, and invasion. A Schematic diagram illustrating the experimental workflow, showing the effects of FGF4 knockdown and overexpression on macrophage secretion of immune suppressive cytokines. B , C ELISA analysis of IL-10 and TGF-β secretion levels from macrophages in the MDA-MB-231 co-culture system. D , E CFSE labeling to assess the effect of macrophages on T cell proliferation, including quantitative analysis. F Experimental workflow diagram showing the effects of M2 macrophage secretions on the proliferation, migration, and invasion of MDA-MB-231 cells. G CCK-8 assay to evaluate the impact of M2 macrophage-conditioned medium on MDA-MB-231 cell proliferation. H Transwell migration and Matrigel invasion assays to assess the effects of M2 macrophage-conditioned medium on MDA-MB-231 cell migration and invasion. (I) Quantitative analysis of migration ability of MDA-MB-231 cells treated with M2 macrophage-conditioned medium, as assessed by Transwell assay. (J) Quantitative analysis of invasion ability of MDA-MB-231 cells treated with M2 macrophage-conditioned medium, as assessed by Transwell assay. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. All cell experiments were performed in triplicate

Article Snippet: After blocking with a blocking solution (Beyotime, China) for 1 h at room temperature, the sections were incubated overnight at 4 °C with primary antibodies against M2 macrophage markers Arg1 (1:200, Cell Signaling Technology, USA, 93668) and CD206 (1:200, BioLegend, USA, C068C2).

Techniques: Migration, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Labeling, CCK-8 Assay, Transwell Assay

Journal: Cell Division

Article Title: FGF4 drives tumor progression in triple-negative breast cancer via IL6/STAT3-mediated macrophage M2 polarization and immune suppression

doi: 10.1186/s13008-025-00164-y

Figure Lengend Snippet: Effects of FGF4 on tumor growth and the immune microenvironment in a TNBC mouse model. A Schematic diagram of the orthotopic tumor model, showing the implantation of FGF4 overexpressing, knockdown, and control breast cancer cells into the mammary fat pads of immunocompetent mice. B Representative images of tumors and tumor weight measurements from each group of mice. C – G In tumor tissues, Immunohistochemistry staining and quantitative analysis of M2 macrophage markers (Arg1, CD206) and M1 macrophage markers (iNOS, CD86). H , I ELISA measurement of immune suppressive cytokine IL-10 levels in tumor tissue. (J-K) Flow cytometry analysis of IFN-γ, TNF-α, and GzmB expression levels in tumor-infiltrating T cells, assessing T cell anti-tumor activity. Data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Each group included 10 mice

Article Snippet: After blocking with a blocking solution (Beyotime, China) for 1 h at room temperature, the sections were incubated overnight at 4 °C with primary antibodies against M2 macrophage markers Arg1 (1:200, Cell Signaling Technology, USA, 93668) and CD206 (1:200, BioLegend, USA, C068C2).

Techniques: Knockdown, Control, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Activity Assay