Figure 10.

IIF analysis of HA-tagged Hic-5 localization in transiently transfected Cos-1 cells that were extracted before fixation. Cos-1 cells transiently transfected with HA-tagged Hic-5 were fixed with paraformaldehyde and processed for IIF analysis with an anti-HA antibody with the use of standard procedures (Nishiya et al., 1999). Before fixation, cells were permeabilized with a low-salt and detergent buffer. Representative cells are shown in A and B. C shows cells that were subsequently digested with DNAse I and extracted with ammonium sulfate. Arrow in B indicates an HA-tagged Hic-5–positive cell devoid of nuclear staining.