FIGURE 2.

Expression and normal cellular distribution are preserved in HTY domain mutant RUNX2 protein. HeLa cells were seeded on gelatin-coated glass coverslips and transfected with 1 μg of HA-tagged wild-type or mutant Runx2 expression plasmid. Cells were processed for in situ immunofluorescence. Whole cells were fixed in 4% formaldehyde, permeabilized, and stained with a polyclonal RUNX2 antibody followed by anti-rabbit Alexa488 secondary antibody. A similar pattern of punctate nuclear signal is observed for all mutant RUNX2 proteins. Nuclei are revealed by 4′,6-diamidino-2-phenylindole staining (DAPI). Differential interference contrast images represent bright field microscopy of respective cells. All magnifications are 63×.