Fig. 6

USP7 controls IRF4 and IRF8 expression by regulating Ezh2. A Crystal structure of overall protein–protein binding between USP7 and Ezh2. B Ribbon representation of the crystal structure of USP7:Ezh2 peptide complex with the USP7 peptide represented by blue color and Ezh2 peptide represented by red color (left). The interactions formed between USP7 and the Ezh2 peptide are shown as yellow dashed lines. The residues involved in the interactions are labeled (right) and presented in table form. C–G Splenic DCs purified from NOD and NOD.Stat5b-CA mice were incubated with either vehicle (0.1% DMSO) or with the Ezh2 inhibitor GSK343 (3 μM/mL) alone or together with the USP7 inhibitor P5091 (5 μM/mL) for 24 h. Thereafter, cells were washed and then stained with anti-CD11c, anti-Ezh2, anti-IRF4, and anti-IRF8 antibodies and analyzed by flow cytometry. C, D Histogram profile (left panel) and Mean fluorescence intensity (MFI) quantification (right panel) of the level of Ezh2 expression in CD11c⁺ DCs. Data are shown as the mean ± SEM of three independent experiments. The two-tailed unpaired Student’s t-test was used. **P < 0.01. E MFI values of Ezh2 in DCs treated with either vehicle or P5091 inhibitor. F, G MFI values of IRF4 and IRF8 in vehicle-treated DCs or DCs treated with Ezh2 inhibitor alone or together with P5091 inhibitor. Data are shown as the mean ± SEM of three independent experiments. One-way ANOVA followed by Tukey’s post hoc test was used. n.s, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001