Figure 3

Effect of fluvastatin and celecoxib on U87- MG cell cycle, proliferation and apoptosis. (a) Cells were treated for 24 hours with 240 μM fluvastatin/100 μM celecoxib. Cell cycle of control and treated cells was analyzed by FACS using propidium-iodide-stained nuclei. Percentage of cells in G1, S and G2 phases is shown. (b) Cells were treated for 24 hours with 240 μM fluvastatin/100 μM Celecoxib. Cell proliferation was analyzed by FACS using KI67 staining. (A) Representative experiment of 3 independent experiments is shown. (B) The percentage of KI67-positive cells with mean plus standard deviation of 3 independent experiments is shown. (c) U87-MG cell lines were cultured with fluvastatin (240 μM) or celecoxib (100 μM) either alone or in combination. After 24 hours, cells were collected and analyzed for fluorescein annexin-V and propidium iodide (PI) labelling by FACS in order to distinguish and quantitatively analyze non-apoptotic cells (Annexin-V negative/PI negative, lower-left), early apoptotic cells (Annexin -V positive/PI negative, lower-right), late apoptotic/necrotic cells (Annexin-V positive/PI positive, upper-right) and dead cells (Annexin V negative/PI positive, upper-left). This double-labelling was performed on untreated cells and treated cells with the drug combination (100 μM of celecoxib/240 μM of fluvastatin).