The C-terminal 95 amino acids of CBFβ-SMMHC are not required for RUNX1 repression, but are important for the deregulation of gene expression. (A) Bar graph from a representative experiment showing fold change in luciferase activity in 293-0 cells transfected with expression plasmids for the indicated proteins. To control for differences in transfection efficiencies, luciferase activity was standardized to renilla activity, which was expressed from a constitutively active promoter. Each transfection was performed in triplicate, and the assay was performed more than 3 times, each time with the same relative luciferase activity. (B) Western blot of 293-0 cells transfected with plasmids expressing the indicated proteins. The blot was probed with an antibody specific to CBFβ, stripped, and reprobed with an antibody specific to tubulin. (C) Venn diagrams representing the numbers of genes that showed > 2-fold change in expression and a P < .05 in the peripheral blood from E12.5 embryos from line no. 2 of the indicated genotypes compared with their wild-type littermate controls. (D) Bar graph of the 10 biologic functions most significantly associated with the differentially expressed genes in Cbfb+/MYH11 embryos, and the P value of these functions for the differentially expressed genes in Cbfb+/ΔC95 and CbfbΔC95/ΔC95 embryos. The vertical line is the statistical significance threshold at the −log of P = .05, therefore functions with P values above this line (to the right) are considered statistically significant.
