Fig 2: T7-MUC1 conjugate induces tumor-specific immune responses. (A and B) Percentages of (A) CD4+/CD3+ T cells and (B) CD8+/CD3+ T cells in the total splenocytes were measured using flow cytometry. (C) Induction of cytolytic T cells in the tumor model mice was measured using a CTL assay. (D) Induction of ADCC in the tumor model mice was measured using an ADCC assay. (E and F) Representative flow cytometry results of CD8+/CD3+ T cells in splenocytes in the (E) PBS and (F) T7-MUC1 groups. Data are presented as the mean ± SD; n=8. *P<0.05 vs. PBS. MUC1, mucin 1; T7, Toll-like receptor 7 agonist; NK, natural killer; ADCC, antibody-dependent cell-mediated cytotoxicity; CTL, cytotoxic T lymphocytes.

Fig 3: Tumor-specific features were conserved in tumoroid models.a–b Paired normal, pleomorphic adenoma (PA), mucoepidermoid carcinoma (MEC), and adenoid cystic carcinoma (AdCC) tissues were processed for organoid cultures. a Organoid growth was observed under a brightfield microscope after 2 weeks of culture in tumor-GEM. Scale bars indicate 100 µm. b Histological examination was performed via H&E staining of tissues (top) and organoids (bottom). Scale bars indicate 50 µm. c The PA-specific expression of PLAG1 was assessed in tissues (left) and organoids (right) using immunohistochemistry (IHC) staining. Paired adjacent tissues with normal histology were used as control. Nuclei were stained with hematoxylin. Scale bars indicate 50 µm. d The AdCC-specific expression of c-Kit was assessed in tissues (left) and organoids (right) using IHC staining. Paired adjacent tissues with normal histology were used as control. Nuclei were stained with hematoxylin. Scale bars indicate 50 µm. e The MEC-specific expression of MUC1 was assessed in tissues (left) and organoids (right) using IHC staining. Paired adjacent tissues with normal histology were used as control. Nuclei were stained with hematoxylin. Scale bars indicate 50 µm. f, g Tumoroids were cultured with or without the p53 inhibitor nutlin-3 (10 µM) for 5 days. Nutlin-3-sensitive (top) or -resistant (bottom) organoids were assessed for growth under a brightfield microscope f while the expression of p53 was evaluated via IHC g. Nuclei were stained with hematoxylin in IHC data. Scale bars indicate 500 µm in (f) and 50 µm in (g). h Diverse sensitivity to radiation was observed in MEC or AdCC tumoroids. Cell viability was assessed at 1, 2, 3, 5, and 10 Gy (n = 5). Cell viability of tumoroids without irradiation was normalized as 100% viability. Data are representative of at least three independent experiments. Source data are provided as a Source Data file.

Fig 4: YBX1 and MUC1 expression was the highly correlated and affected prognosis. (A) YBX1 and MUC1 mRNA expression were based on the Broet Lung and Bhattacharjee Lung datasets from the Oncomine database (p<0.05, p<0.05, respectively). (B) Western blot analysis of YBX1 and MUC1 expression in 6 pairs of lung adenocarcinoma (T) and adjacent tissues (N) (r = 0.5864, P < 0.0037, Red dot: T, Black dot: N). (C) Western blot analysis of YBX1 and MUC1 expression in HLF and 6 lung adenocarcinoma cell lines (r = 0.8789, P < 0.0057). (D) RT-PCR analysis of YBX1 and MUC1 expression in HLF and lung adenocarcinoma cell lines (r = 0.9306, P < 0.0018). (E) IHC analysis of YBX1 and MUC1 expression in lung adenocarcinoma. Original magnification 200× and 400× in the inset. (F) YBX1 and MUC1 expression was positive correlated in 176 lung adenocarcinoma specimens (r = 0.357, P < 0.001). (G) The OS curves of YBX1/MUC1 expression in 176 patients with lung adenocarcinoma (p <0.001). (H) The DFS curves of YBX1/MUC1 expression in 176 patients (p <0.001). The data are presented as mean ± SD of three independent tests.

Fig 5: Threonine attenuates melanogenesis and metastasis‐related gene expression. (a) The human melanoma cell MNT1 was treated with 1–1000 nmol L–1 threonine and melanin levels were visualized. (b) The melanin levels were assessed, and (c) tyrosinase activity was calculated compared with that in the control group. (d–k) The relative expression of melanogenic genes that were altered by threonine treatment is depicted. (l–q) The expression changes in mucin genes are indicated. The data shown are the mean ± SD from three independent experiments. (r–t) The expression levels of the mucin proteins MUCL1, EMCN and MUC1. The data, produced in six independent experiments, are presented as the mean ± SD (pg mL–1). *P < 0·01 and **P < 0·001 compared with the control group.