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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of <t>DAPTA</t> and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.
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PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of DAPTA and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.

Journal: Advanced Science

Article Title: PLAUR + Neutrophils Drive Anti‐PD‐1 Therapy Resistance in Patients with Hepatocellular Carcinoma by Shaping an Immunosuppressive Microenvironment

doi: 10.1002/advs.202507167

Figure Lengend Snippet: PLAUR inhibitor reverses the immunosuppressive phenotype of neutrophils and attenuates tumor progression. A) Flowchart depicting the screening strategy for small‐molecule compounds targeting PLAUR. B) The affinity of 40 candidate small‐molecule compounds targeting PLAUR detected by SPR analysis. C) Schematic diagram of the predicted docking structure of DAPTA and PLAUR. D) Cell viability of DAPTA detected in human neutrophils. E) Western blot analysis of PLAUR, phosphorylated and non‐phosphorylated NF‐κB expression in human neutrophils treated with DMSO or DAPTA (5 µ m ). F) quantitative PCR analysis of the indicated genes in DMSO and DAPTA treatment groups ( n = 3 per group). G) Flow cytometry analysis of CD206 and CD95 expression in DMSO and DAPTA treatment groups (n = 3 per group). H) Schematic showing the treatment plan and establishment of spontaneous HCC models in mice. I) Representative images of the spontaneous tumors at the study endpoint (5 mice per group). Scale bar: 1 cm. J) The maximum tumor volume of each group at the study endpoint ( n = 5 per group). K) Kaplan‐Meier survival curves for mice ( n = 5 per group). L) Immunofluorescence staining and statistical analysis of PLAUR, F4/80 and CD8 in the indicated groups ( n = 5 per group). Scale bars: 100 µm. M,N) Flow cytometry analysis of Ly6G + CD206 + neutrophils, CD11b + F4/80 + macrophages and CD3 + CD8 + T cells in the indicated groups ( n = 5 per group). The data are presented as the means ± SDs. * P < 0.05, ** P < 0.01, and *** P < 0.001, Student's t test.

Article Snippet: For PLAUR inhibitor treatment, mice were treated intraperitoneally with D‐Ala‐peptide T‐amide (DAPTA) (MedChemExpress, 100 μg per mouse) following the schedule shown in Figures and .

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Immunofluorescence, Staining