fig6

Figure 6. Analysis of the specificity of interaction of RUNX2/CD44-ICD deletion constructs. A: immunoblotting analysis with a RUNX2 antibody (top panel): Nuclear lysates prepared from PC3 cells transfected with indicated constructs were immunoprecipitated with a GFP antibody (lane 1-6) and immunoblotted (IB) with an antibody to RUNX2 (lanes 1-6). An equal amount of nuclear lysate used for immunoprecipitation (IP) was assessed by direct immunoblotting of lysates with an antibody to nucleoporin (Input for IP). A decrease in loading was observed in D2 samples, which corresponded to the possible decrease in the coprecipitation of RUNX2; B: confocal microscopy analyses of cells transfected with CD44-ICD deletion construct and stained with an antibody to GFP (green) and RUNX2 (red). Colocalization is seen in yellow (indicated by arrows in FL, D1, D2, D3, and D4). Scale bar: 25 µm. CD44: Cluster of differentiation 44; ICD: intracellular domain