Fig. 3

Autophagy induced by CUDC907 in ESCC cell lines and its regulation in autophagy flux. a Gene function enrichment analysis was performed using the Gene Ontology (GO) database after CUDC-907 treatment in ESCC cell lines. b Formation of autophagosomes showed by cellular fluorescence image. CUDC-907-treated ESCC cells (KYSE-450 and KYSE-510) obtained punctate profile of GFP-LC3B. Scale bar: 20 μm. c Autophagy-related protein expressions of Beclin-1, Atg5, Atg7, LC3B-I/II, and P62 were determined by western blot analysis. d Autophagic flux experiment was verified by western blot analysis using 3-MA (5 mM) or CQ (20 µM) in KYSE-450 cells. e, f Western blot analysis of P62 and LC3B-I/lC3-II. Cell viability and LDH release in CUDC-907 (30 nM, 48 h)-treated KYSE-450 and KYSE-510 cells, pretreated with 3-MA (5 mM) or CQ (20 μM) for 12 h. Error bars are ± SD. *P < 0.05; **P < 0.01; ***P < 0.001