Figure 2

Analysis of differential 5-methylcytidine content of U2OS xenografts using histological sections. Analysis of the relative levels of cellular 5-methylcytidine within xenografts derived from representative control untreated mice (panel A); or decitabine treated mice (panel B) using immunohistochemical staining with 5-mc-Ab. To the right a 20X enlargement of representative U2OS histology is shown. More 5-mc-Ab staining is evident in the nuclei from control sections (panel A-right side) in comparison to nuclear staining in the sections derived from decitabine treated mice (panel B right side). These data are consistent with a reduction in 5-methylcytidine nuclear content in U2OS xenografts derived from mice treated with decitabine. Enlargements of darkly stained host murine kidney cells correspond to heavily methylated differentiated renal tissue (denoted 2 in panels A and B). In panel C the staining intensity from control (dark columns) and decitabine treated (light columns) is quantitated using Aperio scanning image analysis of sections. The graph to the right of panel C confirms that the 5-mc-Ab staining in control untreated (dark columns) U2OS xenograft sections is more intense than the decitabine treated (light columns) U2OS xenografts. This decrease in staining intensity was significant (p < 0.05). In contrast host kidney cells from both control and treated mice do not exhibit any significant difference in staining intensities.