PRAK Interacts with DJ-1 and Prevents Oxidative Stress-Induced Cell Death

Figure 2

Interaction between PRAK and DJ-1 in vitro and in vivo. (a) SDS-PAGE analysis of interaction of His-PRAK with either GST-DJ-1 or GST in vitro. The protein bands are visualized by Coomassie blue staining. (b) The equal input of His-PRAK, GST-DJ-1, and GST on SDS-PAGE. (c) HEK293 cells were cotransfected with pcDNA3/HA-PRAK and pcDNA3/Flag-DJ-1. pcDNA3-Flag was used as the control. Cell lysates were precipitated with anti-flag M2 beads, and both immunoprecipitates (upper) and cell lysates (lower) were immunoblotted with either anti-HA or anti-Flag antibodies. (d) Cell lysates from naive Hela cells were precipitated with protein A/G beads coupled with IgG (left) or anti-PRAK antibody (right). Both immunoprecipitates (upper) and cell lysates (lower) were immunoblotted with either anti-DJ-1 or anti-PRAK antibodies. (e) CFP-DJ-1 and YFP-PRAK were coexpressed in Hela cells followed by observation with different fluorescence channels CFP, YFP, and FRET. The FRET efficiency depicted as a color-coded scale ranging from 0 to 100%. Coexpression of either CFP and YFP-PRAK or YFP and CFP-DJ-1 in Hela cells was used as the control.

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