FIG. 6.

Hdm2 mutants lacking an NrLS in the RING domain remain in the nucleoplasm and sequester ARF within the same compartment. NIH 3T3 cells were transfected with expression plasmids encoding full-length Hdm2 and different Hdm2 mutants as indicated together with GFP-ARF. Hdm2 was detected with antibody 2A10. The numbered squares show nuclear DNA staining by Hoechst dye (top row; blue), GFP-ARF fluorescence (second row; green), Hdm2 fluorescence (third row; red), and Hdm2-ARF overlap (fourth row; yellow). The dark unstained regions within the nuclei correspond to nucleoli, as confirmed by using antibodies to fibrillarin (data not shown) (47).