Figure 2
From: Mitochondrial Thioredoxin System as a Modulator of Cyclophilin D Redox State
Redox Western blot of CypD (A,A′), Trx2 (B) and Prx3 (C) in isolated rat heart mitochondria. Aliquots of 200 μg proteins of rat heart mitochondria were incubated under various conditions at 25 °C for 30 min. (a) control; (b) 2 mM NAC; (c) 2 mM diamide; (d) 1 μM CsA; (e) 5 μM AF; (f) 5 μM AF +1 μM CsA; (g) 2 mM H2O2. To determine the redox state of CypD and Trx2, samples, derivatized first with 30 mM IAA and then with 10 mM IAM, were subjected to urea-PAGE in non-reducing conditions (A,B). To analyze the redox state of Prx3, samples, derivatized with 10 mM AIS, were subjected to SDS-PAGE in non-reducing conditions (C). (A′) densitometric analysis of data reported in panel (A) and referred to CypD and performed using ImageJ software. Bands undergoing oxidation are marked by arrows in (A′).
