fig1

Figure 1. Immunoblotting and confocal microscopy analysis of the expression and distribution of CD44, CD44-ICD and RUNX2 in PCa cell lines. A-C: an equal amount of protein lysates (40 µg) made from LNCaP (lane 1), PC3 (lane 2) and PCa2b (lane 3) cells were immunoblotted with CD44 (A), CD44-ICD (B), and RUNX2 (C) antibodies to detect total cellular levels of the respective proteins. (*) 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. Immunoblotting with a GAPDH antibody was used as a loading control; D: immunostaining analysis of the distribution of RUNX2 (red), CD44-ICD (green), and DAPI (blue). Arrows point to the regions of colocalization (yellow) in RUNX2/CD44-ICD panel. Scale bar: 100 µm. The results represent one of the three separate experiments performed with the same results. CD44: Cluster of differentiation 44; ICD: intracellular domain