Fig. 2. Post-mitotic restoration of the spatially ordered Runx subnuclear organization is functionally conserved. ROS 17/2.8 osteosarcoma cells (top panel) were subjected to in situ immunofluorescence microscopy for endogenous Runx2. Runx2 is distributed at punctate subnuclear domains throughout the interphase and telophase nucleus (lower panel). Subnuclear organization parameters were computed from deconvoluted images for Runx2 for interphase nuclei (I), and both progeny telophase nuclei, denoted at random as telophase nucleus 1 (T₁) or telophase nucleus 2 (T₂). A color map has been applied to the standardized data assigning red to higher values and green to lower values (see supplementary material). Each increment of one reflects one (row) standard deviation (inner left and right panels). ANOVA was performed to assess the significance of observed differences between T₁, T₂, and I nuclei. Asterisks indicate statistically significant differences based on a 0.05 level with correction for false discovery rate. Bonferroni's multiple comparison tests were use to determine which nuclei differed significantly at a 0.05 level. In each case significant differences were observed between telophase (T₁,T₂) and interphase nuclei (I), but differences were not observed between telophase nuclei. Overall mean Clark and Evans statistics (Ro/Re) were 1.4 for Runx2, indicating a non-random organization with spatial order. Bar, 10 µm.