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

Figure 4

Overexpression of PRAK influences the intracellular distribution of DJ-1. (a) NIH3T3 cells were cotransfected with pcDNA3-Flag-DJ-1 and pcDNA3-HA-PRAK. pcDNA3-HA was used as the control. Both cytosolic and nuclear fractions of cell lysates were analyzed by Western blot with anti-Flag or anti-HA antibodies. TATA binding protein (TBP) and β-actin were used as internal controls for nuclear and cytosolic proteins, respectively. (b) The relative intensities of Flag-DJ-1 protein bands from Western blot were analyzed and data are expressed as the mean ± SD of four separate experiments. compared with Flag-DJ-1 in the cytosol fraction from cells transfected with pcDNA3-HA; compared with Flag-DJ-1 in the nuclear fraction from cells transfected with pcDNA3-HA. (c) NIH3T3 cells were cotransfected with pcDNA3-EGFP-DJ-1 and pcDNA3-HA-PRAK or pcDNA3-EGFP-DJ-1 and pcDNA3-HA as the control. Nuclei were stained with DAPI. Scale bar = 20 μm.

(a)

(b)

(c)