Figure 2

Lysine 36 and/or lysine 37 of PARP-2 are required for nuclear localization. A) HEK293T cells were transfected with HA-tagged wild type (wt) PARP-2 or with the indicated mutants. Cells were fixed with methanol for subsequent detection of HA-tagged proteins by immunofluorescence using an anti-HA antibody and a FITC-conjugated anti-mouse antibody. Representative confocal images are presented. B) Lysines 19, 20, 36 and 37 of PARP-2 were changed to glutamic acid or methionine as indicated. C) The nuclear localization is independent of the charge but dependent on the structure of the NLS. As for Fig. 2A, HEK293T cells were transfected with HA-tagged wild type (wt) PARP-2 or with the indicated K → E and K → M mutants and overexpressed proteins were detected as described for Fig. 2A. Representative confocal images are presented.