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. 2011 Apr 25;286(23):20335–20344. doi: 10.1074/jbc.M110.215632

FIGURE 3.

FIGURE 3.

Interaction with PARP-1 is essential for the nuclear localization of KLF8. A, KLF8 mislocalizes in the cytoplasm when its PARP-1 binding site is disrupted or PARP-1 is knocked down but not when PARP-1 activity is inhibited. PARP-1+/+ MEFs were transfected with KLF8 or its PARP-1 binding-deficient mutant (KLF8-mut) individually or in combination with treatment with PARP-1 siRNA (siPARP-1) or inhibitor (PJ-34, 3 μm). GFP siRNA was used as negative control. After 34–48 h, cells were analyzed by anti-HA staining of the KLF8 proteins (green) and Hoechst staining of the nuclei (blue) followed by fluorescent microscopy. Scale bar, 20 μm. B, cytoplasmic mislocalization of KLF8 in PARP-1−/− MEFs can be prevented by ectopic expression of both wild-type PARP-1 and its PARylase dead mutant (E988K). PARP-1−/− cells were transfected with KLF8 or KLF8-mut alone or along with PARP-1 or its dead mutant. After 24 h, cells were analyzed by anti-HA staining of the KLF8 (green), anti-Myc staining of PARP-1, and Hoechst staining of the nuclei (blue) followed by fluorescent microscopy. C and D, statistical analysis of data represented in A and B. The data represent the mean ± S.E. of at least three independent experiments. For each experiment, 200 cells were examined. *, p < 0.01 for difference in nuclear localization (Nuc) only.