FIG. 5.

Phosphorylation of HP1γ is required for recruitment of HP1γ to SAHF but not its localization to PML bodies. (A) HA-tagged wild-type (WT) HP1γ or HP1γ(S93A) was coexpressed with an activated oncogene Ras (H-RasV12) in WI38 cells by retrovirus infection and drug-selected with 3 μg/ml puromycin and 500 μg/ml G418. Eights days postinfection, drug-selected cells were stained with the indicated antibodies and DAPI. Yellow arrows mark HP1γ wild type and HA-HP1γ(S93A) in a PML body, and yellow arrowheads mark HP1γ wild type in SAHF. The latter is largely absent from SAHF. Images were collected by confocal microscopy. (B) One hundred cells shown in panel A were scored for the colocalization of PML and HP1γ wild type or HP1γ(S93A).