Membrane affinity and plasma membrane targeting of GagPol with C-terminal truncations.

<p>(A and B) HeLa cells were transfected with the Gag-FLAG, GagPol-HA, and C-terminally truncated constructs. Gag-FLAG was used as a positive control. (A) Intracellular localization of the truncated GagPol proteins. At 24 hr post-transfection, cells were immunostained with anti-FLAG (red) or anti-HA (green) antibodies and nuclei were stained with TO-PRO-3 (blue). Bottom panels show confocal images overlaid with differential interference contrast images. All micrographs are shown at the same magnification. In each sample, approximately 100 antigen-positive cells (from 3 or 4 independent experiments) were subjected to analysis of the antigen distribution pattern. (B) Membrane affinity of the truncated GagPol proteins. Cells were subjected to membrane flotation centrifugation followed by Western blotting using anti-p24 antibody. Representative blots in three independent experiments were shown.