Fig.3

.Autophagy inhibition significantly reduces the characteristics of OCSCs. (a) 3AO and SKOV3 spheres were seeded at 200 cells per well in a 96-well ultralow attach plates and transfected with lentivirus expressing shRNA for ATG5 (shATG5) or scramble shRNA (Ctrl), or treated with 10 μM CQ (in the last 3 days). Spheres were observed through a microscope (left) and the size and number of the spheres were quantified after 6 days (right). (b) The CD24⁻ phenotype of 3AO spheres (upper) and the CD44⁺ phenotype of SKOV3 spheres (bottom) was determined by FCM after autophagy was inhibited by ATG5 knockdown or CQ(10 μM, 48 h). (c) 3AO and SKOV3 sphere cells were seeded in ultra-low attachment 96-well plates and exposed to paclitaxel at various concentrations for 48 h when autophagy was inhibited by ATG5 knockdown or treated with CQ (10 μM, 48 h). Live cells were measured using the CCK-8 kit. Cell viability was measured by ratio of absorbance value of paclitaxel treated/untreated wells in each group. Three independent experiments were performed and the results were expressed as the means ± SD, and analyzed using Student’s t-test (* P < 0.05, **P < 0.01)