Figure 3.

The protein level of CHIP, its association with SENP3, and its redox state remain unchanged upon oxidative stress. (A) HeLa cells were treated with the indicated concentrations of H₂O₂ for 1 h. SENP3 and CHIP protein levels were evaluated, respectively, by IB. (B) Myc-CHIP and RGS-SENP3 were co-transfected into HEK293T cells for 48 h with MG132 (10 μM) incubation for the later 12 h. H₂O₂ was added at the indicated concentrations for the last 1 h. Exogenous proteins were co-immunoprecipitated using anti-RGS and immunoblotted using the indicated antibodies. (C) HeLa cells were treated with 100 μM H₂O₂ for 1 h. CHIP protein was examined by redox diagonal electrophoresis and IB using CHIP antibody. The blots with short- and long-time exposure were displayed to show the position of CHIP relative to the diagonal line.