Figure 5

Effect of mPARM-1 and hPARM-1 on cell cycle of NIH/3T3 cells. (a) Synchronized NIH/3T3 cells were either untransfected, or transfected with the empty vector (pcDNA3.1A/Myc-His), murine or human Parm-1 constructs. Cells were fixed at 72h post-transfection, stained with propidium iodide and analyzed for cell cycle phase distribution. Percentages of cells in different phases of cell cycle were determined with the ModFit software. (b) Proliferation of control pcDNA3.1A/Myc-His, mParm-1-pcDNA3.1A or hParm-1-pcDNA3.1A transfected NIH/3T3 cells was analyzed using BrdU, 48h after transfection. DAPI labeled nuclei are in purple and cells that have incorporated BrdU are in green. Representative fields were photographed. (c) The percentage of proliferating cells was determined using the following formula: % of proliferating cells = (number of BrdU incorporating cells/total number of DAPI stained cells) * 100. Values were normalized relative to control cells. For a, b and c, similar results were obtained using either PARM-1 tagged GFP or Myc-His (data not shown). All results represent the average of three independent experiments. Statistical analysis was performed using one-way analysis of variance (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001).