Fig. 1: Downregulated TPX2 expression in HCC-infiltrated CD8+ T cells indicated poor antitumor activity.

A Human HCC samples were divided into two groups (TPX2^low/TPX2^high) according to the results of immunohistochemistry (IHC) analysis of TPX2-stained tumor-infiltrating lymphocytes. Immunofluorescent staining of PD-1 and TIM-3 and DAPI staining (for nuclear staining) were performed in the corresponding TPX2^low and TPX2^high HCC samples. Scale bar: 50 μm. B The expression of TPX2 mRNA was detected in CD3+, CD4+, and CD8+ T cells isolated from peripheral blood mononuclear cells (PBMCs) or tumor-infiltrating lymphocytes (TILs). C Expression of the TPX2 protein was detected in CD3+, CD4+, and CD8+ T cells isolated from peripheral blood mononuclear cells (PBMCs) or tumor-infiltrating lymphocytes (TILs). D Tumor-infiltrating CD3+, CD4+, and CD8+ T cells were divided into the TPX2^low and TPX2^high groups according to their expression of TPX2 detected by flow cytometry (FC). The activity (CD69) and proliferation (Ki67) of TPX2^low and TPX2^high CD3+, CD4+, and CD8+ T cells were assayed by FC. The median value of FI (Intensity of fluorescence) was used as the cutoff for the gating strategy for TPX2^high and TPX2^low groups. **p < 0.01, ***p < 0.001; the two-tailed unpaired Student’s t-test was used to compare two groups. PBMCs peripheral blood mononuclear cells, HCC hepatocellular carcinoma, MFI mean fluorescence intensity, TIL tumor-infiltrating lymphocytes. Raw western blot results can be found in the Original data file.