Dominant negative(DN) p73 or p73 silencing reverse the PIM-2 effect on cells over-expressing the 34 kDa PIM-2 isoform.

<p>(<b>A</b>) Light microscopic view (×40) of cells 48 hours after transfecting equal amounts of cells with an empty HA vector as control (control), HA-Pim2 plasmid alone (Pim-2) and a HA-Pim-2 plasmid together with a dominant negative form of p73 (Pim-2+p73DN), and under identical culture conditions. (<b>B</b>) Percent of sub-G1 apoptotic cells, and (<b>C</b>) percent of cells in the G1 phase of the cell cycle, in the specified cultures, as revealed by FACS analysis. Results represent an average of four independent experiments. Asterisk represents statistical significance p<0.01. (<b>D</b>) Western analysis showing siRNA-silencing of p73 compared to untreated control or control of cell treated with scrambled siRNA. (<b>E</b>) FACS analysis of cell cycle pattern of PI stained cells to which siRNAs were introduced [control scrambled (scr.) or p73-directed siRNA (p73)] 24 h prior to transfection with either empty HA plasmid (HA+scr. or HA+73) or HA-Pim-2 plasmid (PIM-2+scr. or PIM-2+73). FACS analysis was performed 76 h after transfection with plasmids. Horizontal line in each pattern indicates the channels that were included in calculation of the sub-G1 phase. (<b>F</b>) Percent of cells at the sub-G1 phase calculated from the cell cycle patterns presented in panel E.