EZH2 overexpression induces proliferation of HPASMCs.

<p>HPASMCs were transfected with a plasmid expressing EZH2 or control plasmids using 4D-Nucleofector system. A) EZH2 protein expression was analyzed by western blot (upper panel) and mRNA expression was analyzed by real-time PCR (lower panel) to determine the transfection efficiency. B) The transfected cells were fixed and labeled with PI after 48 hrs and analyzed for cell cycle by flow cytometry. Percentage number of cells in S and G2/M phase of the cell cycle is plotted (upper panel) and representative histograms from the cell cycle analysis of EZH2 and control GFP plasmid transfected cells are shown (lower panel). C) Real-time PCR analysis of cell proliferation markers expressed as normalized fold expression is shown. All experiments were repeated at least 3 times and graphs shown are average of three independent experiments.