Fig. 4

Inhibition of USP7 promotes cDC1 over cDC2 in tolerogenic Stat5b-CA.DCs. Purified splenic DCs (1 × 10⁵ cells/well) from NOD and NOD.Stat5b-CA mice were cultured in the presence (+ P5091) or absence (− P5091) of USP7 inhibitor (5 µM/mL) for 24 h. DCs were washed, labeled with anti-CD11c mAbs in combination with anti-CD11b, anti-Sirpα, and anti-XCR1 mAbs. A, C, E FACS profiles showing CD11b, XCR1, and Sirpa expression in splenic CD11c⁺ DCs treated with or without USP7 inhibitor. B, D, F Representative bar graphs of CD11c⁺CD11b⁺, CD11b⁺Sirpa⁺, CD11b⁻XCR1⁺ cDC frequencies. (G, I, and K) Representative histograms of CD11b, Sirpα, and XCR1 gated on CD11c⁺ as assessed by flow cytometry. H, J, L Mean fluorescence intensity (MFI) values of CD11b, Sirpα (gated on CD11c⁺CD11b⁺ DCs), and XCR1 (gated on CD11c⁺CD11b⁻ DCs) expression. Data are shown as the mean ± SEM of at least four 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