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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 <t>and</t> <t>CCL5</t> expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) <t>ELISA</t> measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.
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CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 and CCL5 expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) ELISA measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.

Journal: Journal of Advanced Research

Article Title: CCR1 hi /CCL5 hi macrophage-mediated CCL5 hi T cell chemotaxis in salivary gland aggravates Sjögren’s syndrome

doi: 10.1016/j.jare.2025.06.076

Figure Lengend Snippet: CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 and CCL5 expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) ELISA measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.

Article Snippet: The release level of CCL5 in cell culture supernatants was assessed using a commercial ELISA kit (EK1129, Lianke Biotechnology Co., Ltd., Hangzhou, China) following the manufacturer’s instructions.

Techniques: Migration, Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Transfection, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay