Fig. 4. The region responsible for subnuclear targeting of Runx2 resides in the C-terminus. ROS 17/2.8 cells grown on gelatin-coated coverslips were transfected with (A) Runx2 (1-528) (B) Runx2 (1-516) and (C) Runx2 (1-376). The cells were then processed for NM-IF preparation and in situ immunofluorescence. The coverslips were incubated with monoclonal antibody against HA tag (1:3000) followed by incubation with Alexa 568 conjugated secondary antibody (1:800) to detect the expressed proteins. Images were taken using Zeiss Axioplan digital microscope and Metamorph software was used for bio imaging. Bars (2 µm) represent the relative magnification of the images.