Research Paper Volume 12, Issue 2 pp 1656—1684

Figure 5. High LSP1 expression contributed to the immunosuppressive microenvironment in GBM. (A and B) The correlation analysis of LSP1 expression and non-tumor immune and stromal cell populations in GBM by MCP-counter (A, CGGA RNA sequencing dataset, n = 138; B, TCGA RNA sequencing dataset, n = 155; with Pearson correlation analysis). (C and D) Association of LSP1 expression with tumor purity, immune and stromal score, and twenty-four immune cell populations in GBM microenvironment by GSVA. (C, CGGA RNA sequencing dataset, n = 138; D, TCGA RNA sequencing dataset, n = 155; with Pearson correlation analysis). (E and F) Representative IHC images (E, 200X, scale bar = 50μm) and analysis (F) verifying LSP1 expression correlated with macrophages and neutrophil in 29 cases of GBM samples (macrophage: r = 0.4339, P = 0.0187; neutrophil: r = 0.4428, P = 0.0162; n = 15; with Pearson correlation analysis). (G) Representative IF images of LSP1 (red), Neutrophil Elastase (green), and DAPI (blue) staining in clinical GBM samples (n = 3) (200X, scale bar = 50μm). (H) Representative IF images of LSP1 (red), IBA1 (green), and DAPI (blue) staining in clinical GBM samples (n = 3) (200X, scale bar = 50μm). (I) Representative western blot image (left panel) and analysis (right panel) of LSP1 expression in M0 macrophages induced from THP-1 cells. (J) Transwell assay showing LSP1 knockdown inhibit the migration of M0 macrophages, and LSP1 overexpression enhanced their migration. CGGA, Chinese Glioma Genome Atlas; TCGA, The Cancer Genome Atlas; GBM, glioblastoma multiforme; LSP1, lymphocyte specific protein 1; IBA1, ionized calcium binding adapter molecule 1; DAPI, 4’,6-diamidino-2-phenylindole; IDH1, isocitrate dehydrogenase 1.