Osteosarcoma remains an aggressive solid tumor affecting mainly children and youngadults, and despite advances in treatment, a deeper mechanistic understanding of thedisease is still needed. Among the pathways associated with osteosarcoma progression,Platelet-Derived Growth Factor Receptor (PDGFR) downstream signaling has emergedas a potential target. PDGFR and its ligands have been reported to be overexpressed inosteosarcoma, activating multiple downstream signaling cascades involved inproliferation, tumor survival, and cancer progression, although many of thesemechanisms remain incompletely understood.Proteome Integral Solubility Alteration (PISA) combined with LC-MS basedproteomics represents an approach for studying protein thermal stability changesassociated with cellular signaling and protein interaction dynamics. In this study, PISAwas used to investigate proteomic alterations induced downstream of PDGF-BBstimulation in U2OS osteosarcoma cells. Differential thermal stability analysiscombined with STRING, Reactome and CORUM characterization was used to identifypathways and protein networks associated with PDGF signaling.Proteins associated with translation, transcription regulation, and cell cycle progressionshowed altered thermal stability following PDGF-BB stimulation. Specifically, proteinsinvolved in ribosome assembly, translation initiation, extracellular matrix, mitoticspindle regulation, and AP-1 mediated transcription were identified within the affectednetworks. The results suggest that PDGF-BB stimulation induces proteome alterationsassociated with proliferative and translational cellular responses in U2OS.