Fig. 4. Functional impairment of macrophages in Ubr5-null tumors.

a Peritoneal macrophages were isolated and starved overnight before loaded into upper chamber with ID8 cells cultured at the bottom. Migrated cells were assessed after 16 h incubation (n = 5 biologically independent samples per group and an average of five fields acquired from each sample). Scale bars: 200 μm. b Heat map representation of differentially expressed M1/M2 genes in naïve macrophages and TAMs. c Representative FACS analysis of intracellular Arginase-1 in CD45⁺ CD11b⁺ F4/80⁺ cells and quantification expressed as mean florescence intensity (MFI) of Arginase-1 (n = 3 mice per group). d Representative FACS analysis and proportion of PD-L1⁺ cells gated on CD45⁺ CD11b⁺F4/80⁺ cells (n = 3 mice per group). e T cell proliferation suppression assay. CFSE-labeled T cells from naïve mice were stimulated with anti-CD3 and CD28 antibodies and co-cultured for 3 days with TAMs isolated from ID8/GFP or ID8/Ubr5^−/− bearing mice at 3:1 ratio (n = 3 mice per group). f Proportion of Muc16⁺CD45⁻ tumor cells with or without exogenous TAMs on day 45 post-tumor inoculation (n = 3 mice per group). g Kaplan–Meier curves showing the survival of ID8 bearing mice with or without exogenous TAMs (n = 5 mice per group), P = 0.0026, log-rank test. Data are representative of two independent experiments with similar results. Data are presented as mean ± SEM, unpaired two-sided Student’s t-test with no correction for multiple comparison,*P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.