Analysis of the effects of OPN over expression on cell morphology and RANKL expression in PC3 cell lines. A. Indicated PC3 cell lines were photographed using a phase contrast microscope. An increase in multinucleated giant cells was observed in PC3/OPN cells (magnification × 200). B. Immunoblotting analysis of RANKL expression in PC3 cell lines Immunoblotting analysis in protein lysates made from the indicated cell lines was performed with an antibody to RANKL. PC3/OPN cells express greater level of RANKL that PC3 cells. Results shown are a representation of three independent experiments. Bottom panel shows normalization with GAPDH.