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The sensitivity of the U87vIII CHCHD2 WT and KO cells to a panel of cytotoxic agents was determined using the SRB assay according to previously published protocols (31)

Glycerol release was measured under basal and forskolin-stimulated conditions using a colorimetric assay kit (Sigma-Aldrich)

In addition, SHYS reduced the elevated total iron concentration and FTH/FTL ratio in the renal tissues of these mice, indicating relief of the iron overload