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. 2018 Sep 17;2018:7010472. doi: 10.1155/2018/7010472

Figure 3.

Figure 3

Autophagy is critical for ovarian cancer spheroid cells to enter quiescent state. (a) Immunostaining of Ki-67 in A2780 adherent and spheroid cells. A2780 cells were cultured under adherent or spheroid conditions for 48 h. The spheroid cells were trypsinized and seeded back to attach to the plate for 4 h; both the adherent and spheroid cells were fixed, stained with anti-Ki-67 antibody (green), counterstained with DAPI (blue), and observed with a fluorescence microscope. The experiment was repeated twice with similar results. (b) Flow cytometric analysis of G0 percentage of A2780 cells adherent and spheroid cells. A2780 cells were cultured under adherent or spheroid conditions for 48 h, trypsinized, fixed, and stained with propidium iodide and anti-Ki-67 antibody. Cells were subjected to flow cytometry analysis with Beckman Coulter FACS (mean ± SEM, n = 3). (c) Flow cytometric analysis of G0 percentage of Nc or ATG5 shRNA cells adherent and spheroid cells. Cells were subjected to flow cytometry analysis with BD FACS (mean ± SEM, n = 4).