Supplementary material for Choi et al. (July 3, 2001) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.151236498
Fig. 7.
In vivo targeting of Runx2DC to subnuclear sites is compromised. Cells were obtained from calvarial tissue of WT and homozygous DC mutant mice at 17.5 dpc cultured for 2 days on glass coverslips and prepared for in situ immunohistochemistry of WT and Runx2DC proteins. (Top) WT cells (×100) show punctate staining of Runx2 in whole cells (WC), which is retained in the nuclear scaffold/nuclear matrix-intermediate filament (NMIF) preparations. Homozygous (DC/DC) cells (×100) also show distinct punctate foci in WC, but complete absence of Runx2DC in the NMIF preparations. The antibody control (no primary antibody) shows background fluorescence for comparison. (Middle) Phase microscopy shows cellular and nuclear morphology in WC and NMIF. (Bottom) Nuclei were stained with 4',6-diamidino-2-phenylindole (0.5 mg/ml) to verify removal of DNA during the NMIF extraction procedure.