Fig. 9.

Limited excretion of excess RSS causes the accumulation of persulfidated proteins in cardiomyocytes. (A,B) Intracellular and (C,D) extracellular CysSSH concentrations produced by (A,C) primary hepatocytes and (B,D) primary cardiomyocytes prepared from WT mice, following treatment with 100 μM Na₂S₄ in serum-free medium for 1 h. After treatment, cells were washed three times and incubated in serum-free medium, then cells and media were collected at the indicated time points and their CysSSH concentrations quantified using LC-ESI-MS/MS. (E) SDS-PAGE stained with Flamingo fluorescent gel stain analysis of protein-SSnH in primary cardiomyocytes (CM) and hepatocytes (HC) treated with (+) or without (−) 100 μM Na₂S₄ for 1 h and incubated for an additional 3 h in serum-free medium. Data represent the mean ± SEM (n = 3). **p < 0.01.