Figure 2.

NPLOC4⁺ TAMs mediate hepatocellular carcinoma progression by inhibiting CD8⁺ T cells infiltration and cytotoxic function. (A) A tumor model was established by subcutaneous inoculating HEP53.4 mixed with macrophages (transfected with si-NC or si-NPLOC4, at a 2:1 ratio) into C57BL/6 mice. tumor growth was monitored regularly, and the data of tumor volume was represented as mean ± SEM (n = 5 in each group). (B) Flow cytometry of CD8⁺ T cell ratio to CD45⁺ in NPLOC4 knockdown and control groups was expressed as mean ± SEM (n = 5 in each group). (C) Flow cytometry of the exhausted status of CD8⁺ T cells (PD-1⁺ and TIM-3⁺ labeling) in NPLOC4 knockdown and control groups was expressed as mean ± SEM (n = 5 in each group). (D) Flow cytometry of the cytotoxic function of CD8⁺ T cells (IFN-γ labeling) in NPLOC4 knockdown and control groups was expressed as mean ± SEM (n = 5 in each group). (E) The diagram and tumor weight of tumor growth in orthotopic tumor model inoculating HEP53.4 cells in Nploc4^f/f and Nploc4^cKO groups. (F) Representative images of immunofluorescence staining in orthotopic tumor tissues of the Nploc4^f/f and Nploc4^cKO groups. (G) Flow cytometry of CD8⁺ T cell ratio to CD45⁺ in Nploc4^f/f and Nploc4^cKO groups (n = 5 in each group). (H) Flow cytometry of the exhausted status of CD8⁺ T cells (PD-1⁺ and TIM-3⁺ labeling) in Nploc4^f/f and Nploc4^cKO groups (n = 5 in each group). (I) Flow cytometry of the cytotoxic function of CD8⁺ T cells (IFN-γ labeling) in Nploc4^f/f and Nploc4^cKO groups (n = 5 in each group). (J-L) The diagram (J), tumor volume (K) and weight (L) of tumor growth in mouse model inoculating HEP53.4 cells mixed with macrophages (transfected with si-NC or si-NPLOC4, at a 2:1 ratio) and treated with IgG or CD8 antibody. n: number of biological replicates. *p < 0.05; **p < 0.01; ***p < 0.001.