Figure 5. Notch1 induced autophagy in human osteosarcoma cells. (A) Osteosarcoma cells were transiently transfected with GFP-LC3 plasmid for 24 h and then treated with doxycycline (10 μM) for 0, 12, 24, 36, 48 and 60 h. Doxycycline treated cells displayed a punctate pattern of GFP-LC3 expression, which represented formation of autophagosomes. (B) The histogram indicated that autophagy proportion from three separate experiments in different groups. *P<0.05, significantly different compared with the untreated control group. (C) Autophagy-related proteins, LC3 and beclin-1, were analyzed by western blotting. *P<0.05, significantly different compared with the untreated control group. All the experiments were conducted in triplicates.