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. 2023 May 19;14:2859. doi: 10.1038/s41467-023-38605-3

Fig. 6. USP5 inhibition combination with Trametinib or CTLA-4 blockade has an additive effect on suppressing tumor growth.

Fig. 6

a A schematic treatment plan for immunocompetent BALB/c mice bearing subcutaneous CT26 tumors. Mice were subcutaneously implanted with 1.5 × 105 CT26 cells and treated with control vehicle, USP5 inhibitor (EOAI3402143), MEK inhibitor (Trametinib), or combined treatment, respectively. b, c The growth of CT26 tumors in BALB/c mice with indicated treatments (b). Kaplan–Meier survival curves for BALB/c mice bearing CT26 tumors (c). Data were presented as mean ± S.D. Two-way ANOVA (b), log-rank test (c). n = 6 mice per group. d Representative images from immunohistochemical (IHC) staining of PD-1, CD8, and GzmB in CT26 tumors from BALB/c mice treated with indicated reagents. Scale bar: 50 μm. e, f Quantification for PD-1 (e) and CD8 (f) positive cells were counted from mice tumor sections. Each plot represents the mean of three random fields of each mouse tumor section. n = 4 mice per group. g The growth of MC38 tumors in WT or cKO mice with indicated treatment groups. n = 5 mice per group. Data were presented as mean ± S.D. Two-way ANOVA. h–k Quantification of flow cytometry result of PD-1+ (h), IFN-γ+ (i), TNF+ (j), and GzmB+ (k) cells as percentage of CD8+ T cells in MC38 tumors after indicated treatments. n = 5 each group. l, m The growth of CT26 tumors in BALB/c mice with indicated treatments. Tumor volumes of mice treated with control antibody/vehicle (n = 8), EOAI3402143 (n = 8), anti-CTLA-4 monoclonal antibody (n = 9), or combined therapy (n = 9) were measured every 2 days and plotted individually (l). Kaplan–Meier survival curves for BALB/c mice bearing CT26 tumors (m). Data were presented as mean ± S.D. Log-rank test (m). n Quantification of flow cytometry result of PD-1+ as percentage of CD3+ T cells in CT26 tumors after indicated treatments. n = 6 mice each group. o, p Quantification of flow cytometry result of IFN-γ+ (o), TNF+ (p), and GzmB+ (q) cells as the percentage of CD8+ T cells in LLC tumors after indicated treatments. n = 6 mice each group. For e, f, h–k, n–q, data were presented as mean ± S.D. Two-tailed unpaired t-test. Source data are provided as a Source data file.