Figure 5.
No interaction was identified between Prx1 and ASK1. (A) DOK cells were transfected for 48 h with plasmids encoding Prx1 and ASK1, respectively. Cells were stimulated with 5 mM H2O2 for 30 min. Cell lysates were incubated with rabbit monoclonal antibodies against Prxl, precipitated by Protein A/G beads and detected by western blots using rabbit polyclonal antibodies against ASK1. Pre-immune IgG was used as a negative control. Lane 1, blank cell control; lane 2, transfection control; lane 3, Cells transfected with 2 vectors; lane 4, cells transfected with 2 vectors and treated with H2O2; lane 5, positive control. (B) Direct interaction between Prx1 and ASK1 proteins in vitro was investigated by glutathione S-transferase pull-down assay. Prx1, peroxiredoxin 1; ASK1, apoptosis signal-regulating kinase 1; IP, immunoprecipitation; IB, immunoblotting; IgG, goat anti-rabbit HRP-conjugated IgG.