Fig. 8. (A) The NMTS of Runx2 is functionally autonomous. HeLa cells grown on coverslips were transfected with Gal4 or Gal4-NMTS Runx2 and the cells were then processed for NM-IF preparation and in situ immunofluorescence as described. The expression of the proteins was detected by monoclonal antibody against Gal4 DBD (1:1000). (B) The NMTS of Runx2 can target a heterologous protein to the NM-associated Runx2 foci. ROS 17/2.8 cells grown on gelatin-coated coverslips were transfected with Gal4-NMTS Runx2 and were processed for double immuno labeling for endogenous Runx2 and over-expressed Gal4 DBD constructs. (C) The NMTS of Runx2 exhibits mild transactivation. The HeLa cells stably transfected with luciferase reporter carrying five Gal4 binding sites upstream of transcription initiation site were transiently transfected with Gal4 DBD or Gal4-NMTS. Cells were harvested in reporter lysis buffer 24 hours after transfection and assayed for luciferase activity as described.