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: To induce differentiation into macrophages, THP-1 cells were treated with 100 nM PMA for 24 h. Differentiation was confirmed by immunofluorescence analysis of CD68 localization and expression using a mouse anti-CD68 antibody (1:500, 66231–2-Ig, Proteintech, Wuhan, China).
Techniques: Migration, Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Transfection, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay