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f4 80 staining  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc f4 80 staining
    ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b <t>+</t> <t>F4/80</t> + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .
    F4 80 Staining, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1308 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/f4+80+staining/F4%2F80+XP+Rabbit+mAb/pmc12885477-287-12-15
    Average 98 stars, based on 1308 article reviews
    f4 80 staining - by Bioz Stars, 2026-09
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    Images

    1) Product Images from "Citalopram exhibits immune-dependent anti-tumor effects by modulating C5aR1 + TAMs"

    Article Title: Citalopram exhibits immune-dependent anti-tumor effects by modulating C5aR1 + TAMs

    Journal: eLife

    doi: 10.7554/eLife.103016

    ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .
    Figure Legend Snippet: ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .

    Techniques Used: Western Blot, Knockdown, Injection, Immunofluorescence, Isolation, Flow Cytometry, Generated, Cell Function Assay, Expressing

    ( A ) Schematic depicting macrophage blockade with clodronate liposomes. Twelve days before tumor inoculation, C57BL/6 mice were pretreated with clodronate liposomes or phosphate-buffered saline (PBS) liposomes. Subsequently, GLUT1 KD Hepa1-6 cells were subcutaneously injected into the C57BL/6 mice. The effect of citalopram (5 mg/kg) on the tumor burden was evaluated after 18 days of drug treatment. ( B ) Immunofluorescence analysis of F4/80 + macrophages in the liver and tumor tissues of indicated groups. ( C ) In C57BL/6 mice, the effect of citalopram on the GLUT1 KD Hepa1-6 xenograft tumors was measured in the presence of macrophage depletion ( n = 7 per group). ( D ) Immunohistochemical analysis of cleaved caspase-3 (CCS3) and Ki67 in GLUT1 KD Hepa1-6-bearing subcutaneous xenograft tumors, treated with DMSO or 5 mg/kg citalopram ( n = 7 per group). Scale bar, 50 μm. ( E ) In the context of macrophage depletion, measurement of CD8 + T cell function in tumor tissues upon DMSO or citalopram treatment. Values are presented as mean ± SD and compared by the Student’s t test ( C–E ).
    Figure Legend Snippet: ( A ) Schematic depicting macrophage blockade with clodronate liposomes. Twelve days before tumor inoculation, C57BL/6 mice were pretreated with clodronate liposomes or phosphate-buffered saline (PBS) liposomes. Subsequently, GLUT1 KD Hepa1-6 cells were subcutaneously injected into the C57BL/6 mice. The effect of citalopram (5 mg/kg) on the tumor burden was evaluated after 18 days of drug treatment. ( B ) Immunofluorescence analysis of F4/80 + macrophages in the liver and tumor tissues of indicated groups. ( C ) In C57BL/6 mice, the effect of citalopram on the GLUT1 KD Hepa1-6 xenograft tumors was measured in the presence of macrophage depletion ( n = 7 per group). ( D ) Immunohistochemical analysis of cleaved caspase-3 (CCS3) and Ki67 in GLUT1 KD Hepa1-6-bearing subcutaneous xenograft tumors, treated with DMSO or 5 mg/kg citalopram ( n = 7 per group). Scale bar, 50 μm. ( E ) In the context of macrophage depletion, measurement of CD8 + T cell function in tumor tissues upon DMSO or citalopram treatment. Values are presented as mean ± SD and compared by the Student’s t test ( C–E ).

    Techniques Used: Liposomes, Saline, Injection, Immunofluorescence, Immunohistochemical staining, Cell Function Assay

    ( A ) Real-time qPCR revealing the mRNA expression of M1-oriented ( Il6 , Ifnb1 , and Nos2 ) and M2-oriented ( Mrc1 , Il10 , and Arg1 ) markers in isolated macrophages from orthotopic Hepa1-6 tumors ( n = 3 per group). ( B, C ) Gating strategies used for flow cytometry analysis of tumor and splenic lymphocytes. Panel A : Identification of CD4 + T cells, CD8 + T cells, and dendritic cells (DC). Panel B : Identification of B220 + B cells, tumor-associated macrophages (TAMs), and natural killer (NK) cells. ( D ) Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages, CD4 + T cells, CD8 + T cells, B220 + B cells, CD11c + DC cells, and NK1.1 + NK cells in spleen tissues from orthotopic xenograft model, which generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( E ) Real-time qPCR analysis of Glut1 and Glut3 expression in intratumoral CD8 + T cells ( n = 3 per group). In all panels, *p < 0.05, **p < 0.01. Values as mean ± SD and compared by the Student’s t test.
    Figure Legend Snippet: ( A ) Real-time qPCR revealing the mRNA expression of M1-oriented ( Il6 , Ifnb1 , and Nos2 ) and M2-oriented ( Mrc1 , Il10 , and Arg1 ) markers in isolated macrophages from orthotopic Hepa1-6 tumors ( n = 3 per group). ( B, C ) Gating strategies used for flow cytometry analysis of tumor and splenic lymphocytes. Panel A : Identification of CD4 + T cells, CD8 + T cells, and dendritic cells (DC). Panel B : Identification of B220 + B cells, tumor-associated macrophages (TAMs), and natural killer (NK) cells. ( D ) Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages, CD4 + T cells, CD8 + T cells, B220 + B cells, CD11c + DC cells, and NK1.1 + NK cells in spleen tissues from orthotopic xenograft model, which generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( E ) Real-time qPCR analysis of Glut1 and Glut3 expression in intratumoral CD8 + T cells ( n = 3 per group). In all panels, *p < 0.05, **p < 0.01. Values as mean ± SD and compared by the Student’s t test.

    Techniques Used: Expressing, Isolation, Flow Cytometry, Generated

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    ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b <t>+</t> <t>F4/80</t> + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .
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    Proteintech immunofluorescence staining
    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative <t>immunofluorescence</t> merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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    f4/80, inos, arg-1 immunofluorescent (if) staining  (Servicebio Inc)
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    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative <t>immunofluorescence</t> merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative <t>immunofluorescence</t> merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative <t>immunofluorescence</t> merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative <t>immunofluorescence</t> merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
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    Image Search Results


    ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .

    Journal: eLife

    Article Title: Citalopram exhibits immune-dependent anti-tumor effects by modulating C5aR1 + TAMs

    doi: 10.7554/eLife.103016

    Figure Lengend Snippet: ( A ) Western blotting showed the knockdown efficiency of GLUT1 in mouse Hepa1-6 cells. ( B ) GLUT1 KD Hepa1-6 cells were subcutaneously injected into the Rag1 −/− or immunocompetent C57BL/6 mice, and mice were treated with 5 mg/kg citalopram when bore visible tumors; 3 weeks later, tumor burden was examined ( n = 6–7 per group). ( C ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host ( n = 7). ( D ) Immunofluorescence analysis of C5a deposition in GLUT1 KD Hepa1-6 tumors from C5ar1 +/− and C5ar1 −/− C57BL/6 host. Scale bar, 50 μm. ( E ) Experimental design of bone marrow transfer experiments. ( F, G, I ) GLUT1 KD Hepa1-6 cells were subcutaneously implanted into syngeneic recipient (r) mice that had been reconstituted with bone marrow cells from either C5ar1 +/− or C5ar1 −/− donor mice. The therapeutic effect of citalopram ( F ), C5a deposition ( G ), and macrophage phagocytosis ( I ) in this model was analyzed. Scale bar, 50 μm. ( H ) The phagocytic capacity of macrophages isolated from GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host. Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages ( J ), CD206 + TAMs and CD11b + TAMs ( K ), tumor-infiltrating lymphocytes ( L ) in tumor tissues from orthotopic xenograft model, which was generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( M, N ) Measurement of CD8 + T cell function in tumor tissues from the groups mentioned in C and F . ( O ) The growth kinetics of GLUT1 KD Hepa1-6 tumors in C5ar1 +/− and C5ar1 −/− C57BL/6 host upon CD8 + T cell depletion ( n = 7). ( P ) Correlation analysis of C5aR1 expression and immune checkpoint molecules, gene signatures of TAMs, exhausted T cells, and effector Tregs in the TCGA cohort ( n = 371). In all panels, *p < 0.05, **p < 0.01, ***p < 0.001; ns, non-significant. Values are presented as mean ± SD and compared by two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons ( B, C, F, O ), Student’s t test ( H–M ), one-way ANOVA multiple comparisons with Tukey’s method ( B, N ), and the Spearman’s rank correlation methods ( P ). Figure 3—source data 1. Original western blots for , indicating the relevant bands. Figure 3—source data 2. Original files for western blot analysis displayed in .

    Article Snippet: When the mice were sacrificed, liver and tumor tissues were extracted, and F4/80 staining (1:250, Cell Signaling Technology, #70076) was performed to determine the efficiency of macrophage depletion.

    Techniques: Western Blot, Knockdown, Injection, Immunofluorescence, Isolation, Flow Cytometry, Generated, Cell Function Assay, Expressing

    ( A ) Schematic depicting macrophage blockade with clodronate liposomes. Twelve days before tumor inoculation, C57BL/6 mice were pretreated with clodronate liposomes or phosphate-buffered saline (PBS) liposomes. Subsequently, GLUT1 KD Hepa1-6 cells were subcutaneously injected into the C57BL/6 mice. The effect of citalopram (5 mg/kg) on the tumor burden was evaluated after 18 days of drug treatment. ( B ) Immunofluorescence analysis of F4/80 + macrophages in the liver and tumor tissues of indicated groups. ( C ) In C57BL/6 mice, the effect of citalopram on the GLUT1 KD Hepa1-6 xenograft tumors was measured in the presence of macrophage depletion ( n = 7 per group). ( D ) Immunohistochemical analysis of cleaved caspase-3 (CCS3) and Ki67 in GLUT1 KD Hepa1-6-bearing subcutaneous xenograft tumors, treated with DMSO or 5 mg/kg citalopram ( n = 7 per group). Scale bar, 50 μm. ( E ) In the context of macrophage depletion, measurement of CD8 + T cell function in tumor tissues upon DMSO or citalopram treatment. Values are presented as mean ± SD and compared by the Student’s t test ( C–E ).

    Journal: eLife

    Article Title: Citalopram exhibits immune-dependent anti-tumor effects by modulating C5aR1 + TAMs

    doi: 10.7554/eLife.103016

    Figure Lengend Snippet: ( A ) Schematic depicting macrophage blockade with clodronate liposomes. Twelve days before tumor inoculation, C57BL/6 mice were pretreated with clodronate liposomes or phosphate-buffered saline (PBS) liposomes. Subsequently, GLUT1 KD Hepa1-6 cells were subcutaneously injected into the C57BL/6 mice. The effect of citalopram (5 mg/kg) on the tumor burden was evaluated after 18 days of drug treatment. ( B ) Immunofluorescence analysis of F4/80 + macrophages in the liver and tumor tissues of indicated groups. ( C ) In C57BL/6 mice, the effect of citalopram on the GLUT1 KD Hepa1-6 xenograft tumors was measured in the presence of macrophage depletion ( n = 7 per group). ( D ) Immunohistochemical analysis of cleaved caspase-3 (CCS3) and Ki67 in GLUT1 KD Hepa1-6-bearing subcutaneous xenograft tumors, treated with DMSO or 5 mg/kg citalopram ( n = 7 per group). Scale bar, 50 μm. ( E ) In the context of macrophage depletion, measurement of CD8 + T cell function in tumor tissues upon DMSO or citalopram treatment. Values are presented as mean ± SD and compared by the Student’s t test ( C–E ).

    Article Snippet: When the mice were sacrificed, liver and tumor tissues were extracted, and F4/80 staining (1:250, Cell Signaling Technology, #70076) was performed to determine the efficiency of macrophage depletion.

    Techniques: Liposomes, Saline, Injection, Immunofluorescence, Immunohistochemical staining, Cell Function Assay

    ( A ) Real-time qPCR revealing the mRNA expression of M1-oriented ( Il6 , Ifnb1 , and Nos2 ) and M2-oriented ( Mrc1 , Il10 , and Arg1 ) markers in isolated macrophages from orthotopic Hepa1-6 tumors ( n = 3 per group). ( B, C ) Gating strategies used for flow cytometry analysis of tumor and splenic lymphocytes. Panel A : Identification of CD4 + T cells, CD8 + T cells, and dendritic cells (DC). Panel B : Identification of B220 + B cells, tumor-associated macrophages (TAMs), and natural killer (NK) cells. ( D ) Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages, CD4 + T cells, CD8 + T cells, B220 + B cells, CD11c + DC cells, and NK1.1 + NK cells in spleen tissues from orthotopic xenograft model, which generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( E ) Real-time qPCR analysis of Glut1 and Glut3 expression in intratumoral CD8 + T cells ( n = 3 per group). In all panels, *p < 0.05, **p < 0.01. Values as mean ± SD and compared by the Student’s t test.

    Journal: eLife

    Article Title: Citalopram exhibits immune-dependent anti-tumor effects by modulating C5aR1 + TAMs

    doi: 10.7554/eLife.103016

    Figure Lengend Snippet: ( A ) Real-time qPCR revealing the mRNA expression of M1-oriented ( Il6 , Ifnb1 , and Nos2 ) and M2-oriented ( Mrc1 , Il10 , and Arg1 ) markers in isolated macrophages from orthotopic Hepa1-6 tumors ( n = 3 per group). ( B, C ) Gating strategies used for flow cytometry analysis of tumor and splenic lymphocytes. Panel A : Identification of CD4 + T cells, CD8 + T cells, and dendritic cells (DC). Panel B : Identification of B220 + B cells, tumor-associated macrophages (TAMs), and natural killer (NK) cells. ( D ) Flow cytometry showed the infiltration of CD45 + CD11b + F4/80 + macrophages, CD4 + T cells, CD8 + T cells, B220 + B cells, CD11c + DC cells, and NK1.1 + NK cells in spleen tissues from orthotopic xenograft model, which generated in immunocompetent C57BL/6 mice with Hepa1-6 cells ( n = 5 per group). ( E ) Real-time qPCR analysis of Glut1 and Glut3 expression in intratumoral CD8 + T cells ( n = 3 per group). In all panels, *p < 0.05, **p < 0.01. Values as mean ± SD and compared by the Student’s t test.

    Article Snippet: When the mice were sacrificed, liver and tumor tissues were extracted, and F4/80 staining (1:250, Cell Signaling Technology, #70076) was performed to determine the efficiency of macrophage depletion.

    Techniques: Expressing, Isolation, Flow Cytometry, Generated

    In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative immunofluorescence merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Journal: Materials Today Bio

    Article Title: Silicified strontium-curcumin chelated nanospheres mitigate inflammatory vicious cycles to accelerate diabetic wound healing

    doi: 10.1016/j.mtbio.2025.102407

    Figure Lengend Snippet: In vitro experiments assessing the anti-inflammatory and antioxidative properties of Sr-Cur NPs. (a) Representative images of intracellular ROS levels in HUVEC and Raw264.7 cells, as indicated by the DCFH-DA probe Scale bar: 100 μm for HUVEC and 50 μm for Raw264.7 cells. (b, c) ROS levels quantitative analysis of the results (n = 3). (d, f, g) Representative immunofluorescence merged images of Raw264.7 cells showing iNOS (green) and CD206 (red) Scale bar: 50 μm, accompanied by quantitative analysis (n = 3). (e, h, i) Protein expression of iNOS and CD206 in Raw264.7 cells and corresponding quantitative analysis (n = 3). (j, k) Assessment of CD206 expression by flow cytometry and its quantification in LPS-treated RAW 264.7 macrophages (n = 3). Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Article Snippet: Immunofluorescence staining for F4/80 (CST, 70076, 1:200), IL-1β (CST, 12242, 1:200), and CD206 (Abcam, ab64693, 1:2000) was used to evaluate inflammation at the wound site.

    Techniques: In Vitro, Immunofluorescence, Expressing, Flow Cytometry, Standard Deviation, Control

    In vivo assessment of regulation of Sr-Cur@Alg on macrophage polarization and its pro-angiogenesis effect. (a) Representative images showing CD206 + (red) and F4/80 + (green) macrophages stained by immunofluorescence. (b) Quantitative analysis of the proportion of CD206 + cells (n = 3). (c) IL-1β representative immunofluorescence image. (d) IL-1β quantitative fluorescence analysis (n = 3). (e) CD31 (red) and α-SMA (green) representative immunofluorescence images. (f) CD31 and α-SMA quantitative analysis (n = 3). (g) iNOS, TNF-α, Arg-1, and CD206 protein expression on day 7 post-Sr-Cur@Alg treatment. (h–k) Quantitative protein analysis (n = 3) All Scale bar: 100 μm. Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Journal: Materials Today Bio

    Article Title: Silicified strontium-curcumin chelated nanospheres mitigate inflammatory vicious cycles to accelerate diabetic wound healing

    doi: 10.1016/j.mtbio.2025.102407

    Figure Lengend Snippet: In vivo assessment of regulation of Sr-Cur@Alg on macrophage polarization and its pro-angiogenesis effect. (a) Representative images showing CD206 + (red) and F4/80 + (green) macrophages stained by immunofluorescence. (b) Quantitative analysis of the proportion of CD206 + cells (n = 3). (c) IL-1β representative immunofluorescence image. (d) IL-1β quantitative fluorescence analysis (n = 3). (e) CD31 (red) and α-SMA (green) representative immunofluorescence images. (f) CD31 and α-SMA quantitative analysis (n = 3). (g) iNOS, TNF-α, Arg-1, and CD206 protein expression on day 7 post-Sr-Cur@Alg treatment. (h–k) Quantitative protein analysis (n = 3) All Scale bar: 100 μm. Data are expressed as mean ± standard deviation, and one-way analysis of variance (one-way ANOVA) was used for statistical comparisons. All experimental groups were compared to the PBS control group (∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Article Snippet: Immunofluorescence staining for F4/80 (CST, 70076, 1:200), IL-1β (CST, 12242, 1:200), and CD206 (Abcam, ab64693, 1:2000) was used to evaluate inflammation at the wound site.

    Techniques: In Vivo, Staining, Immunofluorescence, Fluorescence, Expressing, Standard Deviation, Control