Glioblastoma multiforme (GBM) remains the most aggressive and challenging central nervous system tumor due to the heterogeneity of the tumor microenvironment, prompting suboptimal effects to immune checkpoint blockade treatments.1,2 Notably, biomolecular condensates formed via liquid–liquid phase separation (LLPS) have been implicated in cancer progression by altering the tumor microenvironment and enhancing drug resistance.3, 4, 5 Consequently, it is urgent and imperative to identify valuable differentially expressed LLPS-related genes (DELRGs) and establish a prognostic model for GBM based on LLPS to address the immunosuppressive tumor microenvironment in GBM. Therefore, our study is dedicated to elucidating the machinery of LLPS-related genes in GBM by analyzing transcriptomic and single-cell RNA sequencing data, supported by experimental validation. The workflow for the transcriptomic, clinical, and single-cell RNA sequencing data analysis of GBM patients based on LLPS-related genes is shown in Figure 1.