Fig. 5

Blockade of USP7 downregulates IRF4 and upregulates IRF8 expression. Purified splenic cDCs (1 × 10⁵ cells/well) from NOD and NOD.Stat5b-CA mice were cultured for 24 h in the presence (+ P5091) or absence (− P5091) of USP7 inhibitor (5 µM/mL). cDCs were washed, labeled with anti-CD11c mAbs in combination with Abs anti-IRF-4 or anti-IRF-8, and analyzed by FACS. A, C Counter plots (left) and representative bar graphs (right) showing CD11c⁺IRF4⁺ (A) and CD11c⁺IRF8⁺ (C) cell frequencies in purified splenic cDCs. B, D Flow cytometry histogram showing IRF4 (B) and IRF8 expressions (D) in splenic CD11c⁺ DCs. E, G Flow cytometry histograms showing IRF4 (E) and IRF8 (G) expression in purified splenic CD11c⁺ DCs treated with or without USP7 inhibitor (5 µM/mL) for 24 h. F, H Mean fluorescence intensity (MFI) values of IRF4 (F) and IRF8 (H) expression. Data are shown as the mean ± SEM of five independent experiments. The significance was calculated using One-way ANOVA with Tukey’s post-test. n.s, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001