Fig 1: BSGLWE restricts autophagic flux of OS cells in vitro. (A) Morphological changes induced by BSGLWE were observed by transmission electron microscopy. BSGLWE group cells were treated with BSGLWE (4ng/mL). The yellow arrows mean apoptotic bodies and nuclear condensation. The red arrows mean autophagosomes. Scale bar, 2 μm. (B) The HOS and U2 osteosarcoma cells were treated with different concentrations of BSGLWE for 24 h. The expression of those autophagy-related proteins, which included ATG5, ATG12, LC3B, P62, was measured by a Western blot assay. (C, D) An immunofluorescence assay revealed the level of protein LC3B increased in the BSGLWE (4mg/mL) group compared with the NC group in both HOS and U2 cell lines. (E) HOS and U2 cells were treated with different concentrations of BSGLWE (0,4,8 mg/mL), with or without 3-MA (3mM), (F) or with or without CQ (8uM) for 24 h. The protein expressions of LC3B and p62 were measured by Western blotting. (G) HOS and U2 cells were treated with different concentrations of BSGLWE (0,4,6,8 mg/mL), with or without 3-MA (3mM), CCK-8 assay was used to test cell viability. Each assay was repeated 3 times. Magnification, ×200 (C). Scale bar, 100 μm (C). Error bar = mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 versus control, #P< 0.05 versus BSGLWE treatment.