Fig. 2
From: TriPer, an optical probe tuned to the endoplasmic reticulum tracks changes in luminal H2O2
TriPer’s responsiveness to H2O2 in vitro. a–c Traces of time-dependent changes to the redox-sensitive excitation ratio of HyPer (a), TriPer (b), or the TriPer mutant lacking its peroxidatic cysteine (TriPer C199S) (c) in response to increasing concentrations of H2O2 or the reducing agent dithiothreitol (DTT). d–g Non-reducing and reducing SDS-PAGE of recombinant TriPer (d), HyPer (e), a TriPer mutant lacking its peroxidatic cysteine (TriPer C199S) (f), and HyPerC199S along HyPer/TriPer lacking their resolving cysteine (HyPer C208S/TriPer C208S) (g) performed following the incubation in vitro with increasing concentrations of H2O2 for 15 min, black arrow denotes the high molecular weight species, exclusive to TriPer and to HyPer/TriPer lacking their resolving cysteine (HyPerC208S/TriPerC208S), emerging as a result of H2O2 induced dithiol(s) formation in trans. Shown are representatives of n ≥ 3