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. 2013 Jan 29;8(1):e54750. doi: 10.1371/journal.pone.0054750

Figure 3. H2O2 modifies Cys thiols on α-ENaC.

Figure 3

A) Right panel: α-ENaC protein was heterologously expressed (with β- and γ- subunits) and immunoprecipitated from HEK cells. Left panel: Fluoroscein-5-Maleimide (F5M) labeling of free Cys thiols decreases with oxidizing conditions; excitation emission 480/535 nm; non-reducing gel (−DTT). B) Simple linear regression graph of F5M relative light units (RLU) and H2O2 treatment shows relationship between Cys modification and H2O2 concentration. Dependent variable: F5M RLU. Independent variable: mM H2O2. n = 15; p = 0.05. C) Specificity of F5M label. Right lane = mouse lung homogenate labeled with 0.6 mM F5M. Left lane = mouse lung homogenate labeled with 0.6 mM F5M with/without 6 mM glutathionne. D) F5M RLU normalized to β-actin expression.