Fig. 1

VCP impairs CD8⁺T cells infiltration, activation, and effector function to promote HCC progression. a, b Representative images of tumor volumes from Hepa1-6-bearing nude and C57BL/6 mice sacrificed on day 31. The average sizes of tumors were measured and plotted (n = 5/group). Mean values ± SEM. Two-way ANOVA. c Schematic illustrating transposon- and CRISPR-Cas9-based injection of vectors into mice via the tail vein to generate liver tumors resembling human HCC. The transposon-based vector overexpressing Myc can express luciferase. Schematic created with BioRender.com. d, e Bioluminescence images of C57BL/6 mice at indicated time points after plasmid injection. Bioluminescence signal at indicated time points post plasmid injection in C57BL/6 mice. f Representative images of livers from (d). g Kaplan–Meier survival curve of C57BL/6 mice treated as in (d). Log-rank test. h, i t-SNE analysis of scRNA-seq data of 4 subclusters from CD8⁺T cells. j, k Representative images of tumor volumes from Hepa1-6-bearing C57BL/6 mice treated with anti-CD8 and sacrificed on day 25 (n = 5/group). Mean values ± SEM. Two-way ANOVA. l–n The percentage of CD8⁺T cells (l), cytokines production (Granzyme B, IFN-γ) (m), and activation indicators (CD69, CD25, CD44) (n) in spontaneous tumor were measured by flow cytometry (n = 5/group). o Representative tissue microarray (TMA) images for VCP, CD8a, and GzmB immunohistochemistry in 90 HCC patients. p The IHC staining was scored, and correlation analyses were performed (n = 90 patients). The Pearson correlation test was used. q Signature score of VCP and CD8A expression in patient #3 and #4. r Patients clinical samples were stained with multiple immunofluorescence for DAPI, CD8a, GzmB, and VCP. Data are presented as mean values ± SD. Statistical significance was determined using two-sided t-tests, *P < 0.05, **P < 0.01, and ***P < 0.001