fig3

Figure 3. Analysis of CD44-ICD overexpression and its interaction with RUNX2. A: an equal amount of protein lysates (40 μg) prepared from PC3 cells transfected with CD44-ICD or control PC3 cells were used for immunoblotting analysis with a CD44-ICD antibody. Immunoblotting with a GAPDH antibody was used as a loading control; B: equal amounts of PC3 lysates (200 µg) were immunoprecipitated with a RUNX2 antibody (lane 2-3) or a species-specific non-immune serum (NI, lane 1). Immunoprecipitates were subjected to immunoblotting with an antibody to CD44-ICD. (*) and (**) represent the ~20-kDa and ~25-kDa fragments of CD44 extracellular truncation fragment (CD44-EXT). CD44-ICD is ~16.5-kDa fragment of CD44. An equal amount of lysate (Input) used for immunoprecipitation was assessed by direct immunoblotting of lysates with an antibody to nucleoporin. CD44: Cluster of differentiation 44; ICD: intracellular domain