Expression profile of PC3 holoclones, paraclones, parental cells and holoclone-derived tumors. qPCR analysis showed this profile of markers was associated with the holoclones. FAM65B was strongly up regulated in holoclones compared to paraclones, whereas MFI2, LEF1 and IL18R1 were down regulated in holoclones compared to paraclones (panels A-D). RNA samples were those used for microarray analysis. Total RNA was prepared from each of four sequential cell passages for each PC3-derived clone, and from four passages of parental PC3 cells, 48 h after seeding the cells at 105 cells per well of a 6-well plate. Data are shown for 2 passages of each clone as pairs of adjacent bars. These correspond to passages 5 and 7 after the initial isolation of PC3 holoclones. E-H, the high expression of FAM65B in holoclone 2G7 cells was markedly suppressed in the holoclone 2G7-derived tumors (E), whereas MFI2, LEF1 and IL18R1 were expressed at a low level in the holoclone cells and were induced in the resultant tumors (F-H). The bars in the figure represent mean ± SD values based on triplicate analysis.