Research Paper Volume 8, Issue 12 pp 3375—3389

Impaired energy metabolism of senescent muscle satellite cells is associated with oxidative modifications of glycolytic enzymes

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Figure 3. Identification and data mining of modified proteins. Cellular protein extracts from young (30 CPD) and senescent human myoblasts were separated by two-dimensional gel electrophoresis. After the second dimension, gels were either stained with colloidal Coomassie Brilliant Blue G (bottom panels) or electrotransferred onto nitrocellulose membranes for subsequently detection of: carbonylated proteins using the OxyBlotTM kit (A); glycoxidation protein adducts (B) and HNE-modified proteins (C). Presented results are from one representative experiment of three independent experiments using three different batches of cells. Numbers refer to the spots evidenced as consistently increased in senescent cells identified by MS/MS. (D) Venn diagram depicting the distribution of proteins in relation with the modifications studied (see also Table S1, Table S2 and Table S3). (E) Modified proteins were grouped into functional categories through the use of Ingenuity Pathways Analysis. The bars represent the biological functions identified, named in the x-axis. The dotted line represents the threshold above which there are statistically significantly more proteins in a biological function than expected by chance. The identified proteins associated with each pathway are indicated.