Figure 1

H2228 and AsPC-1 cells were induced to a reversible quiescent state. (a) Schematic representation of the cell culture conditions. (b) Proliferation of H2228 cells under GG and HSS conditions. Proliferation rates (left) and representative fluorescent micrographs (right) are shown. The living cells were fluorescently labeled with CellTracker Green CMFDA. (c) Proliferation of H2228 cells under GG and HSS conditions at different glucose concentrations. Proliferation rates are shown. (d) Cell cycle phase of H2228 under GG and HSS conditions. Representative fluorescent micrographs of H2228/fluorescent ubiquitination-based cell cycle indicator (FUCCI) cells (top) and cell cycle distribution (bottom) are shown. (e) Cell cycle analysis by quantitation of DNA content of H2228, AsPC-1, and A549 under GG and HSS conditions. Representative flowcytometric histograms after DNA staining with propidium iodide are shown. (f) Proliferation of AsPC-1 and A549 cells under GG and HSS conditions. Proliferation rates are shown. (a, b, c, d, e, f) GG, general growth conditions; HSS 0.8, hypoxic and serum-starved (1% O₂ and 0.8% FBS) conditions; HSS 0.4, hypoxic and serum-starved (1% O₂ and 0.4% FBS) conditions; HSS, hypoxic and serum-starved (1% O₂ and 0.1% FBS) conditions; HSS-GG, GG conditions after HSS conditions; FBS, fetal bovine serum. **p < 0.01 (versus GG), *p < 0.05 (versus GG). Scale bar: 500 μm, n = 3.