The expression level of MYO1F is correlated with the efficacy of ICB therapy in both humans and mice. (A) Left: MYO1F and MYO1E gene expression of tumor tissue pooled from responder (N = 49) and nonresponder (N = 42) of PRJEB23709_anti–CTLA-4 + anti–PD-1 datasets. Right: The corresponding Kaplan–Meier survival curves. Data are from the TIGER database. (B) Left: MYO1F and MYO1E gene expression pooled from liver hepatocellular carcinoma (LIHC) tumor tissues (N = 369) and normal tissues (N = 160). Right: MYO1F and MYO1E gene expression pooled from 18 types of tumor tissues and normal tissues from the TCGA database via the GEPAI2 website. Gene level in normal tissue was normalized to 1. The 18 cancers are: adrenocortical carcinoma (ACC), bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), colon adenocarcinoma (COAD), lymphoid neoplasm diffuse large B cell lymphoma (DLBC), kidney chromophobe (KICH), LIHC, lung adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), ovarian serous cystadenocarcinoma (OV), pheochromocytoma and paraganglioma (PCPG), prostate adenocarcinoma (PRAD), rectum adenocarcinoma (READ), thyroid carcinoma (THCA), thymoma (THYM), uterine corpus endometrial carcinoma (UCEC), and uterine carcinosarcoma (UCS). (C) Hazard ratio with high and low expression levels of MYO1F in tumor tissue from 18 types of cancers; ratio in low MYO1F tissues was normalized to 1. The data are from TCGA database via the GEPAI2 website. (D) Kaplan–Meier survival curves of sarcoma. SARC, sarcoma. The data are from the TCGA database via the GEPAI2 website, N (high) = 131; N (low) = 131. (E) Left: Immunofluorescence staining of MYO1F on human normal and melanoma tissue. DAPI used for cell nucleus indication. Right: Statistical value of MYO1F fluorescence intensity, normal, N = 5; tumor, N = 45. Scale bar: 500 µm. (F) Left: Tumor tissues were collected. Right: Tumor growth curve over time following subcutaneous injection of 2 × 10⁵ WT and shRNA-Myo1f B16F10 cells into immunocompetent C57BL/6 mice (N = 5). (G) Tumour growth curve over time after inoculation. Immunocompetent C57BL/6 mice were subcutaneously injected with 2 × 10⁵ WT and shRNA-Myo1f B16F10 cells, following with administered intraperitoneally of anti-PD-1 (10 mg/kg)/ anti-CTLA-4 (10 mg/kg) antibodies on days 10, 14, and 18 (N = 10 in each group). After the treatment with antibodies, tumors were extracted and photographed. (H) Left: Tumor tissues were collected. WT and Myo1f^−/− C57BL/6 mice were subcutaneously injected with 2 × 10⁵ B16F10 cells. Anti–PD-1 (10 mg/kg) and anti–CTLA-4 (10 mg/kg) were administered intraperitoneally on days 10, 14, and 18 (N = 5 in each group). Right: The corresponding tumor growth over time. Data in F–H represent one experiment of three independent repeats, and E represents one experiment of two independent repeats. Data are presented as mean ± SD. P values were analyzed by one-way ANOVA test (F–H); nonparametric Wilcoxon matched-pairs signed-rank test (A and B); Kaplan–Meier (D); and two-tailed unpaired Student’s t test (C and E); *P < 0.05, **P < 0.01, and ***P < 0.001. ns, no significance.