Fig 2: PROS1 increased proliferation, migration, and invasion of Glioma cells in vitro. (A) Single-cell analysis showed the functional state of PROS1 in Glioma. (B) CancerSEA analysis demonstrated that PROS1 positively correlated with invasion. (C, D) The western blotting and qRT-PCR were applied to analyze the protein and mRNA expression level of PROS1 after transfection by siPROS1 or PROS1 knockdown control for 24 h in LN-229 or U-87MG cells. Wound healing assays (scale bar: 500 µm) (E) and Transwell assays (scale bar: 100 µm) (F) were performed in transfected LN-229 or U-87MG cells to evaluate cell migration ability. (G) Cell proliferation of LN-229 or U-87MG cells after knocking down was determined by CCK8 assays. (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).

Fig 3: The relationship of immune cell infiltration and tumor immune microenvironment with PROS1 expression in GBM and LGG. (A) The relationship between PROS1 expression and the immune cell infiltration analyzed by ssGSEA in glioma. The PROS1 expression levels had a significant correlation with the infiltration of (B) pDCs, (C) NK CD56 Bright cells, (D) Macrophages, and (E) Neutrophils in glioma. The correlation of PROS1 expression level with (F) EstimateScore, (G) ImmuneScore, and (H) StromalScore in the high PROS1 and low PROS1 group in glioma. (*, P < 0.05; **, P < 0.01; ***, P < 0.001).

Fig 4: PROS1 expression is significantly correlated with the markers of immunosuppressive molecules in glioma. Enrichment plots from gene set enrichment analysis (GSEA) showed the high expression of PROS1 associated with (A) T cell antigen receptor pathway, (B) PD-1 signaling pathway, and (C) immunotherapy by PD1 blockade. (D) The Spearman correlation analysis revealed that the expression of PROS1 was correlated with the T cell exhaustion, including PD-L1, PD-1, PDCD1LG2, CTLA4, HAVCR2, and GZMB in glioma (all P < 0.05). (E) The Spearman correlation analysis results suggested the immune marker sets of MDSC (CD33, ITGAM, FUT4), TAM (CCL2, CD68, IL-10) and, Treg (FOXP3, CCR8, TGFß1) were associated with the PROS1 expression in glioma.

Fig 5: PROS1 expression correlates with marker sets of immune cells in GBM and LGG. (A–D) The correlation of PROS1 expression with Macrophages, M1 macrophages, and M2 macrophages infiltration level in glioma from the TIMER2.0 database. Scatter plots illustrate correlations between PROS1 expression and markers of M1 (IRF5, NOS2 and PTGS2) (E, F), and M2 macrophages (CD163, MS4A4A, VSIG4) (G, H) in GBM and LGG.