Figure 5: Degradation of redox-sensitive proteins under oxidative stress. | Scientific Reports

Figure 5: Degradation of redox-sensitive proteins under oxidative stress.

From: Degradation of Redox-Sensitive Proteins including Peroxiredoxins and DJ-1 is Promoted by Oxidation-induced Conformational Changes and Ubiquitination

Figure 5

(a) 2D-gel images of Prxs and DJ-1 in MDA-MB-231 cells during recovery with CHX after H2O2 treatment. MDA-MB-231 cells treated with 0.5 mM H2O2 in HBSS for 1 h, were washed, and incubated for 12 h in EMEM supplemented with 10% FBS, with CHX (25 μg/mL). (b,c) Degradation kinetics of Prxs and DJ-1 were confirmed by western analysis. MDA-MB-231 cells treated with various concentrations of H2O2 (0 mM (○), 0.25 mM (▲), 0.5 mM (■)), were incubated in EMEM supplemented with 10% FBS, with CHX (25 μg/mL) for indicated time. Cell lysates were separated on SDS-PAGE and detected with western analysis using various antibodies (b). Quantified data of (b) were presented as graph (c). *p < 0.05; **p < 0.01 versus control. (d) Degradations of oxidized Prx1, 2, 6 and DJ-1 were blocked by proteasome inhibitor lactacystin and autophagy inhibitor 3-MA. MDA-MB-231 cells treated with 0.5 mM H2O2 in HBSS for 1 h were incubated for 15 h in EMEM supplemented with 10% FBS, CHX (25 μg/mL) and lactacystin (1 and 5 μm), an irreversible proteasome inhibitor, and 3-MA (1 and 5 mM), an autophagy inhibitor.

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