Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-2

With Leptomycin B (LMB) to inhibit nuclear export, followed by immunofluorescence as described in Fig. 2. Representative images are presented.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p

Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-3

Blot using a monoclonal anti-HA antibody. 50 μg of whole cell extracts were used, endogenous PARP-1 levels served as loading control. HEK293T cells were transfected with HA-tagged wild type (wt) PARP-2 or with the PARP-2 mutants K36R and K37R. HA-tagged proteins were detected by immunofluorescence as described for Figure 2A. Representative images are presented.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p

Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-1

Ubsequent detection of HA-tagged proteins by immunofluorescence using an anti-HA antibody and a FITC-conjugated anti-mouse antibody. Representative confocal images are presented. Lysines 19, 20, 36 and 37 of PARP-2 were changed to glutamic acid or methionine as indicated. 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.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p

Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-5

Ignments were performed using ClustalW2. Schematic illustration of PARP-2 K → R mutant proteins used in this study: K19, K20, K36 and K37 were changed to arginine using site-directed mutagenesis. Double and quadruple mutants were generated. HA-tagged wild type (wt) PARP-2 or the indicated double or quadruple mutants were expressed in HEK293T cells and expression was analyzed by western blot using a monoclonal anti-HA antibody. 100 μg of whole cell extracts were used, endogenous PARP-1 levels served as loading control.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p

Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-6

Ubsequent detection of HA-tagged proteins by immunofluorescence using an anti-HA antibody and a FITC-conjugated anti-mouse antibody. Representative confocal images are presented. Lysines 19, 20, 36 and 37 of PARP-2 were changed to glutamic acid or methionine as indicated. 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.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p

Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS-0

Ignments were performed using ClustalW2. Schematic illustration of PARP-2 K → R mutant proteins used in this study: K19, K20, K36 and K37 were changed to arginine using site-directed mutagenesis. Double and quadruple mutants were generated. HA-tagged wild type (wt) PARP-2 or the indicated double or quadruple mutants were expressed in HEK293T cells and expression was analyzed by western blot using a monoclonal anti-HA antibody. 100 μg of whole cell extracts were used, endogenous PARP-1 levels served as loading control.<p><b>Copyright information:</b></p><p>Taken from "Importin alpha binding and nuclear localization of PARP-2 is dependent on lysine 36, which is located within a predicted classical NLS"</p><p>http://www.biomedcentral.com/1471-2121/9/39</p><p>BMC Cell Biology 2008;9():39-39.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2496901.</p><p></p