Immunohistochemistry on TMA derived from normal and cancerous prostate tissue. Immunohistochemical staining was performed in prostate cancer and normal tissue microarray with an antibody to RANKL (A and B), RUNX2 (C and D), Smad 5 (E and F) and p-Smad 5 (G-H). Normal tissue adjacent to prostate cancer are shown in A, C, E and G. Prostate carcinoma at grade 2–3 are shown in B, D, F and H. Arrows in C’, D’ G’ and H’ point to nuclear localization of RUNX2 and p-Smad-5 proteins. An arrow in A’ points to a prostate carcinoma filled lumen (A’) adjacent to normal tissue (indicated by an asterisk; A’). Sections were immunostained (brown) with indicated primary antibody as described in the Methods section. Immunostained sections were counterstained with hematoxylin stain (blue). Magnification is 50X in A-H. Location of the high magnification (X200) regions shown in A’-H’ is indicated by a rectangle field in A-H. Staining was repeated two times.