Fig. 2

TSSC3 overexpression enhances autophagy flux in osteosarcoma cells in vitro. a MTF and SaOS2 cells were stably transfected as indicated. Representative electron microscopic images showing that an increased number of autophagic vacuoles (AVs; autophagosomes and autolysosomes) are present in MTF and SaoS2 cells transfected with lenti-overTSSC3. The numbers of AVs in each cell were quantified by randomly selecting 20 cells per group. *P < 0.05, Scale bars: 1 μm. b Representative images of the western blotting analysis of autophagy-related proteins levels in MTF and SaOS2 cells. Cells were treated with or without TSSC3-overexpression. Protein levels on the western blots were quantified by densitometry of TSSC3, ATG5, BECN1, LC3-II, and P62, normalized to GAPDH and shown as a fold change compared with that of the control (TSSC3-) groups. Quantification data was in Additional file 3: Figure S2d. c Western blotting of LC3-II and p62 in cells following TSSC3-overexpression or treatment with autophagy flux inhibitors CQ for 12 h as indicated. Protein levels on the western blots were quantified by densitometry of LC3-II and P62, normalized to GAPDH, and shown as a fold change compared with that of the control (TSSC3- and CQ-) groups