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Fig. 4. Discrimination between wild-type Runx2 and NMTS mutants on the basis of domain size, packing, and spatial randomness. To understand the subnuclear organization of the wild-type Runx protein and the five mutants, we analyzed factors scores, which reflect the sum of standardized subnuclear organization parameters multiplied by respective factor loadings. Factor scores assign a value to each of the unobservable factors (factor A: domain size properties, factor B: domain packing, and factor C: domain spatial randomness). Using the data acquired from the 330 nuclear image sections, we computed factor scores for wild-type and each of the mutants and analyzed star-plots of these scores on three axes (see supplementary material). The center of the star-plot has a value of -0.5, the end of each axis has a value of 0.5, and the mid-point on each axis is zero; these values are in standardized units. The three mean factor scores for each protein define the points of a filled triangle that has been drawn to illustrate the similarities and differences among each of the proteins. Based upon the shape of each of the filled triangles, we can discriminate two groups of domain organizations: one comprised of the wild-type Runx2 protein along with the Y407A and R398A mutants and a second group containing Y433A, Y428A, and the functionally compromised Runx2-
C mutant. Differences in the shape of the triangles highlight the selective alterations in subnuclear organization as a consequence of NMTS mutations.
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