Fig. 2

Aurora-B knockdown induces autophagy in OS. The expression of LC3 protein in tumor tissues were detected by immunohistochemical staining (a), and analyzed by staining scores was performed in 69 OS patients. There are 37 cases of low Aurora-B expression and 32 cases of high Aurora-B expression. b *P < 0.05.143B and HOS cells were infected with shAurora-B and scrambled lentivirus. Aurora-B expression in 143B and HOS cells was knocked down using shAurora-B lentivirus, and analyzed by western blotting (c) and qRT-PCR (d). ****P < 0.01. The RFP-GFP-LC3 fusion assay was performed and cells were visualized by laser scanning confocal microscopy (e); fusion numbers were quantified by randomly selecting three views per slide. ****P < 0.01, Scale bars: 20 μm. Autophagosomes and autolysosomes were observed with a transmission electron microscope (f). Scale bars: 5 μm (left pictures) and 1 μm (right pictures). The protein level of LC3-I, LC3-II, and SQSTM1 were analyzed by western blotting (g). Infected cells were treated with 10 μM CQ (chloroquine diphosphate) for 24 h. The total level of LC3-I, LC3-II, and SQSTM1 proteins were used to detect the autophagic flux (h)