Fig 2: The MYC-USP10-SOX4 axis is upregulated in thymocytes developmental process and T-ALL.a Venn diagram depicting the intersections of predicted results from five datasets. b GEPIA-based analysis GTEx, TCGA (pan-cancer cohorts), and hematopoietic malignancies. Pearson correlation plots are shown. c–d ChIP–PCR assays were performed in HEK293T cells to assess MYC binding to the USP10 promoter (c). Quantification of ChIP enrichment relative to input is shown (n = 1 independent experiment) (d). e Public T-ALL transcriptomic data (GSE26713) show higher MYC and SOX4 expression in T-ALL (n = 117) than in HC (n = 7). Violin plots show distributions with median and interquartile range (dashed). P values: two-sided Mann–Whitney test. f RT-qPCR analysis of MYC, USP10, and SOX4 mRNA levels in PBMCs (n = 11). g IB analysis of USP10 (n = 10) and SOX4 (n = 8) protein levels in PBMCs. h Schematic of experimental design. i Flow cytometry analysis indicating DN-to-DP transition in Rag1⁻/⁻ mice after αCD3ε antibody injection. j IB analysis of c-Myc and USP10 protein levels following treatment (n = 3). k RT-qPCR analysis of Usp10 mRNA expression in DN3 thymocytes (n = 5). l IB analysis of USP10 and SOX4 protein levels in purified DN3 thymocytes (n = 3). m IB analysis of c-Myc, USP10 and SOX4 expression in thymi (n = 3). n–o DN cells from MycTg/Tg control and MycTg/TgLckcre/+ mice were transduced with control or Usp10 shRNA, cultured 3 days, and analyzed for CD4, CD8 and Ki-67 by flow cytometry (n). Bar plot shows Ki-67⁺ DN cells (n = 8) (o). Representative data from ≥2 independent experiments are shown. n denotes biologically independent samples (or independent experiments with similar results, as indicated). ns, not significant; DN, double-negative; DP, double-positive; SP, single-positive; HC, healthy control; T-ALL, T cell acute lymphoblastic leukemia; RQ, relative quantitation; shNT, non-targeting shRNA; PBMCs, peripheral blood mononuclear cells. P values are shown in the graphs. Statistics: unpaired two-tailed Student’s t test (k) or one-way ANOVA (o). Data are mean ± SEM. Source data are provided.

Fig 3: The role of TLR4 in maintaining trained immunity of ILC2s in the mouse type 2 inflammatory model.A Experimental protocol employing HDM-sensitized Tlr4⁻/⁻ mice. i.n., intranasal administration; HDM, house dust mite. B, C Representative contour plots analysis of total ILC2s (B) and IL-13+ILC2s (C). D Representative image of MUC5AC immunohistochemistry staining picture to evaluate mucus. E Percentage of total cells in the bronchoalveolar lavage fluid (BALF), percentage of total ILC2s and IL-13+ILC2s in lung tissues, concentration of IL-13 in lung tissues, and the percentage of MUC5AC+cells among the epithelial cells of mice (wild-type group, n = 6 mice; Tlr4⁻/⁻group, n = 5 mice). F Experimental protocol employing HDM-sensitized Rag1⁻/⁻ mice treated with TLR4 antibody. i.n., intranasal administration; i.p., intraperitoneally injection; HDM, house dust mite; Ab, antibody. G, H Representative contour plots analysis of total ILC2s (G) and IL-13+ILC2s (H). I Representative image of MUC5AC immunohistochemistry staining picture to evaluate mucus. J Percentage of total cells in the bronchoalveolar lavage fluid (BALF), percentage of total ILC2s and IL-13+ILC2s in lung tissues, concentration of IL-13 in lung tissues, and the percentage of MUC5AC+cells among the epithelial cells of mice (control group, n = 6 mice; anti-TLR4 group, n = 6 mice). K Experimental protocol employing Rag2⁻/⁻Il2rg⁻/⁻C57BL/6 J mice. i.n., intranasal administration; i.v., tail intravenous injection; i.p., intraperitoneally injection; HDM, house dust mite; Ab, antibody. L–O Representative contour plots analysis of eosinophils in lung tissues (L), representative image of MUC5AC immunohistochemistry staining (M), percentage of total cells in the bronchoalveolar lavage fluid (BALF), eosinophils in lung tissues, the percentage of MUC5AC+cells among the epithelial cells of mice (N), and airway hyperreactivity (O) in Rag2⁻/⁻Il2rg⁻/⁻C57BL/6 J mice injected with purified TLR4+ILC2s from HDM-sensitized wild type C57BL/6 J mice and then treated with TLR4 antibody before the following HDM challenge (n = 5 mice in each group). Each dot indicates one mouse, and bars represent mean ± SEM. Data for E and J were pooled from two independent experiments. Results for N to O were confirmed in two independent experimental replicates, with representative data shown. P-values calculated by a two-tailed unpaired t test.

Fig 4: Loss of MBTPS1 inhibits tumor growth in immunocompetent mice and sensitizes tumors to immunotherapy.a Growth curves and weights of shVec/shMbtps1 MC38 tumors in female C57 mice (6-8 weeks old). b, c Growth curves of shVec/shMbtps1 CT26 tumors in female BALB/c mice (6-8 weeks old) and E0771 tumors in C57 mice. d, e Growth curves of OEVec/OEMbtps1 MC38 (d) and E0771 (e) tumors in C57 mice. f, g Growth curves of shVec/shMbtps1 MC38 tumors in female Rag1-/- mice (5-6 weeks old) (f) and female BALB/c Nude mice (5-6 weeks old) (g). h Schematic of in vivo competition assay (scale bar: 100 μm). i, j Ratio changes of E0771 (i) and MC38 (j) cells cultured in vitro or tansplanted in C57 mice. k, l Growth curves (k), weights and tumor image (l, scale bar: 1 cm) of shVec/shMbtps1 MC38 tumors in anti-PD-1 (5 mg/kg, i.p.)-treated C57 mice. m Survival of shVec/shMbtps1 MC38 tumor-bearing mice treated with anti-PD-1 (5 mg/kg, i.p.). n Growth curves of E0771 tumors in anti-PD-1 (5 mg/kg, i.p.)-treated C57 mice. o, p Growth curves of KPC (o) and B16 (p) tumors in anti-PD-1 (10 mg/kg, i.p.)-treated C57 mice. q, r Growth curves (q) and weights (r) of MC38 tumors in C57 mice treated with half-dose anti-PD-1 (2.5 mg/kg). s Survival of E0771-bearing mice with half-dose anti-PD-1 (2.5 mg/kg). n = 6 mice per group in a–g and n–r; n = 8 mice per group in i–l and s; n = 10 mice per group in m. Data are presented as means ± SDs. Two-tailed unpaired Student’s t-test for a–g, i and j. one-way ANOVA with Sidak’s multiple comparisons test for k, l and n–r; Kaplan-Meier analysis with the log-rank test for m and s. Source data are provided as a Source Data file.

Fig 5: The production of cathelicidin (CRAMP in mice and LL‐37 in humans) in mice and in human subjects. Related to Fig 1 Serum CRAMP determination by ELISA. Rag1 −/−/gluten‐free: unsensitized Rag1 −/−mice fed with gluten‐free diet. Rag1 −/−/gluten: unsensitized Rag1 −/− mice fed with gluten‐containing diet (n = 8).Western blot of pro‐form CRAMP and the mature peptide in duodenum of Rag1 −/−, Cnlp −/− Rag1 −/−, C57BL/6 and BALB/c mice.Serum LL‐37 determination by ELISA. Anti‐tTG IgA: anti‐tissue transglutaminase immunoglobulin A antibodies. Anti‐DGP IgG: anti‐deamidated gliadin peptides immunoglobulin G antibodies (Anti‐tTG IgA−Anti‐DGP IgG−, n = 50; Anti‐tTG IgA+Anti‐DGP IgG−, n = 46; Anti‐tTG IgA−Anti‐DGP IgG+, n = 50; Anti‐tTG IgA+Anti‐DGP IgG+, n = 13). Data information: Data (A, C) were representative and were the mean ± SD from three independent experiments. Data in B were representative from three independent experiments. P values were calculated by unpaired two‐tailed t‐test for comparison of two groups (A) or one‐way ANOVA followed by Tukey’s post hoc test for multiple comparisons (C). Source data are available online for this figure.