Figure 3
Nucleolin depletion leads to decreased cell growth and G2/M accumulation. A. Growth curves of HeLa cells, untransfected or transfected with control siRNA # 1 or the siRNA mix #2 and #4. Cells were counted at the indicated time after transfection. The graph represents the average of 5 independent experiments. B. Analysis of cell cycle in nucleolin depleted cells. Untransfected and transfected HeLa cells were stained with propidium iodide and processed for FACS analysis. Histograms from a representative experiment showing G1 and G2/M populations shaded in red and S phase population in blue. Sub G1 represent apoptotic cell population. The vertical and the horizontal axes represent respectively cell number and DNA fluorescence intensity. The percentages of cells in individual cell cycle phases were quantified with ModFit LD software. C. Mitotic index of HeLa cells and human primary fibroblasts transfected with control siRNA or the siRNA mix #2 and #4. After 4 days of transfection, cell populations were stained with an anti phospho S28-H3 antibody and cyclin B1 to detect mitotic cells. Positive cells were scored by microscopic observation. This graph represents the average of the different experiments (DAPI, PH3S28 and cyclin B1) shown in additional file 5.