Supplemental Figure S1
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Fig. S1. Positive and negative controls for the RUBY assay. (A,C,E,G,I) Fluorescence images; (B,D,F,H,J) merged fluorescence/phase-contrast images. Positive control data showing an interaction between the DsRed-tagged Runt domain of Runx2, RUBY-wtRunx2-RD and the CBFβ RUBY fusion protein, RUBY-CBFβ (E,F). Direct interaction between Runx2 and CBFβ has been previously reported (Kagoshima et al., 1993). A mutated form of the Runx2 Runt domain, G151R, was used as a negative control as this mutant has been previously reported not to interact with Runx2 (Akamatsu et al., 1997); cells expressing this mutant fusion protein, RUBY-G151RRunx2-RD, exhibit no fluorescence when cotransfected with the RUBY-CBFβ construct (I,J). HEp-2 cells transfected with (A,B) RUBYwtRunx2-RD alone, (C,D) RUBY-CBFβ alone, (E,F) RUBY-wtRunx2-RD and RUBY-CBFβ, (G,H) RUBY-G151RRunx2-RD alone, (I,J) RUBY-G151RRunx2-RD and RUBY-CBFβ.
