Figure 1

Effects of hypoxia on viability, proliferation, cell-cycle distribution and apoptosis of Kasumi-1 cells. Cells were incubated in normoxia (diamond, NORMO) or hypoxia (square, HYPO) for the indicated times (days). (a) Viable cells were subjected to the trypan blue exclusion test and counted in a hemocytometer. (b) Mitotic cycle phase distribution of cells was measured by flow cytometry following propidium iodide staining; the percentages of cells in the G0/G1 (white), S (light gray) or G2/M (dark gray) phases are reported. (c) The percentage of apoptotic cells was measured by the Annexin V test and flow cytometry. (a–c) Values represent the average±S.E.M. of data from three (a, b) or four (c) independent experiments. (b, c) Differences, as determined by the Student’s t-test for paired samples, were significant at day 3 (indicated in the graphs) and at all later incubation times (*P<0.05; **P<0.01); significance in (b) refers to either G0/G1 or S values. (d) Cell lysates were subjected to SDS-PAGE and immunoblotting with antibodies raised against ETO or the indicated proteins. Equalization of protein loading was verified on the same membrane. Representative results from one out of three independent experiments are shown