Figure 7.

PL-tagged protein transport into the nucleus efficiently. (A) Wild-type and read-through versions of NQO1 were transfected into 293T, and their localization in the nucleus (Nucl) was analyzed as detailed in the Experimental section. Proteasome was inhibited using MG132 where indicated. Anti-FLAG antibody was used to detect NQO1. FLAG* indicates an over-exposed blot. GAPDH and lamin B1 were used as a loading control for total lysate and nucleus, respectively. (B) Isolated nuclei were incubated in a RIPA buffer without (−Det) or with detergents 1% Triton X-100, 0.1% deoxycholate and 0.1% SDS (+Det) on ice for 15 min and then centrifuged at 10000g for 5 min. Supernatant and pellet were analyzed by Western blotting with anti-FLAG or anti-Histone 2B antibodies. A single representative out of four experiments is shown.