Figure 7
PGBD5 partitions between nucleus and cytoplasm in the Daudi Burkitt’s lymphoma B-cell line. Nuclear and cytoplasmic fractions were prepared as described in Methods. (Upper panel) Whole cell, cytoplasmic, and nuclear fractions of 2×105 cells/lane were resolved by 7% SDS-PAGE, blotted, and probed with anti-rPGBD5 antibody. Mouse whole brain extract and human rPGBD5 served as controls. PGBD5 is indicated by an asterisk. Immunoreactive proteins larger than PGBD5 do not immunoprecipitate with anti-rPGBD5 antibody (compare with Figure 8). These proteins may cross-react weakly with antibody after denaturation and immobilization on the membrane, just as the antibody cross-reacts weakly with the hexahistidine tag but not the HA tag on western blotting (data not shown). (Lower panels) Daudi whole cell, cytoplasmic, and nuclear fractions were also probed for lamin A and β-actin to confirm successful partition.