Redox States of Protein Cysteines in Pathways of Protein Turnover and Cytoskeleton Dynamics Are Changed with Aging and Reversed by Slc7a11 Restoration in Mouse Lung Fibroblasts

Figure 1

Schematic flow chart of labeling redox-sensitive cysteines in peptides from young, old, and old Slc7a11-overexpressing primary mouse lung fibroblasts using iodoTMT reagents. Three replicate cultures of young, old, and old fibroblasts transfected with Slc7a11 were plated in separate dishes. Proteins from each dish were labeled sequentially with 2 different iodoTMT tags. For multiplexed mass spectrometry analysis, 3 mixes were produced by pooling labeled proteins from each experimental condition. Thus, each mix contained proteins labeled with 6 different iodoTMT tags. The detailed procedure is described in the main text.