Fig. 7: Immunological changes induced by PD-1 blockade in Nedd8-deficient tumors.

a Six hundred thousand control (ctrl) or Nedd8 knock-out (KO) EO771 cells were injected subcutaneously (s.c.) in 100 μl medium in 6–10 weeks old female C57BL/6NTac mice. On day 7, 10 or 13, 50 μg of an αPD-1 antibody (RMP1-14) or the Rat IgG2a isotype control (2A3) were injected intraperitoneally (i.p.) in 100 μl PBS (7 or 8 mice per group). b Tumor volumes were recorded until day 15, when cells were harvested for analysis using flow cytometry (6 tumors per group). Data were shown as mean ± SEM and tested using unpaired two-tailed T-tests. Percentages of (c) CD25+ T cells, (d) TNFα+ T cells, (e) CD11b+ myeloid cells or (f) macrophages were compared among groups. Each dot represented an individual tumor and the average values were shown, unpaired two-tailed T-test. Tumors were harvested from an independent in vivo study with the same design and mRNA samples were isolated from tumors and quantified using a Nanostring immuno-oncology panel. Differentially expressed genes were shown when comparing (g) Nedd8 KO and control tumors treated with the isotype control antibody, or (h) treated with the PD-1 blockade using a cut-off of Log2 fold changes > 0.5 and p values < 0.05, unpaired two-tailed T-test. Source data are provided as a source data file.