This project investigates the role of α7 nicotinic acetylcholine receptor (α7 nAChR) signaling in glioblastoma using the U87MG human glioblastoma cell line. Cells were treated with selective α7 nAChR ligands (10 μM of agonist PNU282987, 1 μM of positive allosteric modulator PNU120596, 1 μM of antagonist α-cobratoxin) and their combination. Whole-cell lysates were processed for total proteome analysis, proteins were digested in-solution with trypsin and peptides analyzed by nano-ESI-LC-MS/MS on a Thermo Orbitrap Elite using DDA. Data were searched with PEAKS Studio 11 against the UniProtKB Human database and quantified by label-free quantification (LFQ). Proteomic changes in cytoskeletal, metabolic and ER-associated proteins were detected.