Fig. 4: IL7R modulates macrophage polarization and migration in ovarian cancer.

A Schematic of the in vitro co-culture system between bone marrow-derived macrophages (BMDMs) and tumor cells. Images sourced from Figdraw; B qPCR analysis of macrophage polarization markers in BMDMs co-cultured with Il7r-KO murine ovarian cancer cells versus WT controls (N = 3); C flow cytometry quantification of macrophage polarization in BMDMs co-cultured with Il7r-KO or WT tumor cells (N = 3); D Transwell migration assay assessing chemotaxis of BMDMs toward Il7r-KO or WT tumor cells (N = 3); E qPCR analysis of macrophage polarization markers after adding recombinant IL7 protein to WT murine ovarian cancer cells (N = 3); F flow cytometry analysis of macrophage polarization in human ovarian cancer cells treated with exogenous recombinant IL7 (N = 3); G 3D bioprinted co-culture of M0-polarized THP-1 cells with OVCAR3 cells for 5 days, followed by multiplex immunofluorescence to assess the impact of IL7R-knockdown OVCAR3 cells on macrophage polarization (CD68: green; CD206: red; nuclei/DAPI: blue) (N = 3). Scale bar: 20 µm; H multicolor immunofluorescence was performed to verify the expression of tumor-associated macrophage markers in ovarian cancer tissues from mice injected with Il7r-WT and Il7r-KO cells (CD68: green; CD206: red; nuclei/DAPI: blue) (N = 3). Scale bar: 20 µm; I volcano plot of differentially expressed genes in TAMs from Il7r-KO versus WT murine ovarian tumors (single-cell RNA sequencing); J flow cytometry analysis of CD206⁺ TAM proportions in ascites and spleens of mice bearing Il7r-WT or Il7r-KO ID8 tumors (N = 3). Statistical significance: ns (not significant), *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data represent mean ± SEM.