Fig. 1

Schematic describing the Glioma primary cell biomarker Study workflow. Sample preparation A: surgically-removed tissue samples were enzymatically dissociated to single cells and cultured. Control pooled primary cells and grade 4 glioma pooled primary cells were lysed, digested, and labeled with TMT reagent 126 and 130, respectively. TMT-labeled control and grade 4 glioma peptides were mixed and subjected to HPLC fractionation. Candidate screening B: Fractionations obtained (n = 12) were subjected to LC–MS/MS, and the acquired data were analyzed via Proteome Discoverer to obtain differentially expressed proteins in glioma primary cells. Ingenuity pathway analysis (IPA) were performed to further understand the biological significance of the differentially expressed proteins. Validation C: MRM assays for the differentially expressed proteins were developed using synthetic peptides for each protein