Cell cycle profiles of cultured tumor cells treated with etoposide. HeLa/Fucci2 cells (A, B, and C) and NMuMG/Fucci2 cells (D, E, and F) were treated with 0 μM, 1 μM, and 10 μM etoposide (left, middle, and right of each panel, respectively) for 24 h. (A and D) Typical fluorescence images of cells acquired by LCV100 are shown. Scale bar, 10 μm. (D) The red nuclei with chromosome fragmentation and 4C DNA content are labeled with asterisks and open circles, respectively. (B and E) Cell cycle profiles obtained by FACS. Cell cycle phases highlighted by fluorescence are colored. For measuring DNA content, cells were stained with Hoechst33342. Solid and open arrows indicate the 2C and 4C DNA containing cells, respectively. (C and F) Cell cycle profiles (two-dimensional scatter plots and gallery images) obtained by CELAVIEW. For measuring DNA content, cells were stained with DAPI. High-magnification images of the gallery are presented in Figure 4A (HeLa/Fucci2, 0 μM etoposide, for Figure 3C left), Figure 4B (HeLa/Fucci2, 1 μM etoposide, for Figure 3C middle), Figure 4C (HeLa/Fucci2, 10 μM etoposide, for Figure 3C right), Figure 5A (NMuMG/Fucci2, 0 μM etoposide, for Figure 3F left), Figure 5B (NMuMG/Fucci2, 1 μM etoposide, for Figure 3F middle), and Figure 5C (NMuMG/Fucci2, 10 μM etoposide, for Figure 3F right).