Research Paper Volume 12, Issue 2 pp 1656—1684

Elevated lymphocyte specific protein 1 expression is involved in the regulation of leukocyte migration and immunosuppressive microenvironment in glioblastoma

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.