Proteomic Analysis of Hypoxia-Induced Senescence of Human Bone Marrow Mesenchymal Stem Cells

Figure 1

The complete experimental process. Hypoxia in hBMSCs was induced for 0, 4, and 12 hours, and cellular senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining. TMT quantitative proteomic analysis with LC-MS/MS analysis was performed to identify and quantify proteins. The general characterization of enriched proteins was performed by GO, KEGG, and PPI network analyses. PRM analysis was used to validate the candidate proteins (CCND1, UQCRH, and COX7C) with changes in expressions. Cell apoptosis was evaluated using Annexin V/7-AAD staining by flow cytometry. ROS was detected by the fluorescent probe DCFH-DA. Verifications of signaling pathways were evaluated by western blotting.