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. Author manuscript; available in PMC: 2011 May 1.
Published in final edited form as: Prostate. 2010 May 1;70(6):675–688. doi: 10.1002/pros.21112

Figure 3. Immunocytochemistry confirms functionality of overexpressed EZH2 and of shEZH2.

Figure 3

LNCaP, 22Rv1, PC3, and DU145 were grown and fixed on coverslips, then stained with an anti-Histone 3 Lysine 27 Tri-Methylation antibody (α-H3K27Me3) to detect catalytic activity of EZH2 within the cells. GFP Control: Cells infected with the GFP Control lentivirus; EZH2: Cells infected with the EZH2-overexpession lentivirus; shLuc: Cells infected with a control shRNA lentivirus; shEZH2: Cells infected with shEZH2 lentivirus. Overlay panel allows direct comparison of α-H3K27Me3 staining in transduced (GFP positive) versus non-transduced (GFP negative) cells. In all cell lines tested, cells overexpressing EZH2 have more intense staining with α-H3K27Me3, indicating more histone methylation. Cells with EZH2 knockdown lost nearly all staining with α-H3K27Me3, indicating no histone methylation at this position. Scale Bar is equal to 25 Microns. Nuclear staining intensity for H3K27Me3 was quantified and is shown in Table I.