Fig. 8.

Cell cycle analysis of tumor cells isolated from control and C5a expressing tumors demonstrates increased apoptosis and a block in cell cycle progression at early time points. At the indicated times, freshly isolated cells from control and C5a-expressing tumors were stained with propidium iodide (Panels A, B and C) or annexin V (Panel D) and the percentage of cells with different DNA content was calculated. a Cells with <2N DNA content, indicative of apoptotic cells. b Representative histograms showing the DNA content of cells from tumors harvested at 19 days (control tumor cells, left side; cells from EMT6/C5aA1 injected mice, right side). Numbers indicate the cells with, 2N DNA content. c Cells with >2N DNA content, indicative of cells that are proceeding through the cell cycle. d Percent annexin positive cells from tumors harvested at the indicated days. Cells were gated to be propidium iodide negative, annexin positive. Symbols, * P<0.05; ** P<0.01; *** P<0.001, Students t-Test. Hatched bars, control tumors; solid bars, C5a tumors. All experiments were repeated three times with four mice in each group