Figure 7.

Alkylating agents differ in their ability to preserve protein persulfides in cells. A) Concept of using a comparative “switch assay” to visualize protein persulfides in cellular lysates. MBB trapped persulfides can be fluorescently labeled after reduction, while NEM trapped persulfides are destroyed. In contrast, the labeling of conventional protein disulfide bonds does not depend on the choice of alkylating agent. B) Representative gel visualizing fluorescence (upper left panel) or Coomassie staining (lower left panel). Quantification of overall fluorescence in each lane (right panel). Fluorescence intensity was normalized to the Coomassie stain intensity, n=3. Error bars represent SD. ** P≤0.01, based on a two‐tailed unpaired t‐test.