Effects of ectopically expressed miR-101 on the growth, survivability and invasiveness of PC-3 cells. A. Effect of ectopic miR-101 on histone H3-K27 methylation. PC-3 cells were transfected with miR-cont or miR-101 (120 nM). Aliquots of transfected cells were analyzed by Western blots using the indicated antibodies. Relative protein expression is indicated at the bottom of each image. B. Effects of Ezh2 downregulation on proliferation and colony formation of PC-3 cells. In the top and middle panels, aliquots of transfected PC-3 cells in "A" were studied by WST-1 proliferation assay (top) and clonogenic assay (middle). In the bottom panel, PC-3 cells infected by lentivirus carrying the control and Ezh2 siRNAs were tested by WST-1 assay and Ezh2 knockdown tested by Western blot was indicated. C. Effects of ectopic miR-101 on the invasiveness of PC-3 cells using aliquots of transfected PC-3 cells in A. The asterisk indicates p < 0.05 and representative images are presented. D and E. Effects of restored Ezh2 expression on miR-101 transfected PC-3 cells. In D, PC-3 cells were either infected with lentivirus generated from pSL4 vector or pSL4-Ezh2 as indicated. At 48 h post infection, cells were transfected with 120 nM of miR-cont or miR-101 as labeled. After another 48 h, cells were analyzed by Western blots using the indicated antibodies. In E, aliquots of the infected/transfected PC-3 cells with the corresponding sample numbers (1, 2 and 3) in D were studied by invasion assay. Percent invasion is shown with "*" indicating p < 0.05 and representative images.