Fig. 4

LCN2 inhibition regulating CUDC-907 induced autophagic cell death in ESCC cell lines via ROS production. a A volcano plot of the RNA-seq analysis shows that the expression of different genes changed after CUDC-907 treatment. b LCN2 protein expression was assayed using western blot analysis in KYSE-450 and KYSE-510 cells treated with DMSO, 10 or 30 nM CUDC-907. c Flow cytometry for the ROS generation assay in KYSE-450 cells treated with DMSO, 10 nM or 30 nM CUDC-907, or pretreated with NAC + 30 nM CUDC-907. d, e Western blot analysis of Beclin1 and LC3B-I/LC3B-II. Cell viability and LDH release in CUDC-907 (30 nM, 48 h)-treated KYSE-450 and KYSE-510 cells, pretreated with NAC (5 mM) or no treatment for 6 h. f, g Western blot analysis of Beclin1, LCN2, and LC3B-I/lC3-II. Cell viability and LDH release in CUDC-907 (30 nM, 48 h)-treated KYSE-450 and KYSE-510 cells, transfected with siLCN2 or negative control treatment for 48 h. h Flow cytometry as a measure of ROS levels in different types of treatment in KYSE-450 cells, histograms indicate the change in the mean of fluorescence in different groups of KYSE-450 cells. Error bars are ± SD. *P < 0.05; **P < 0.01; ***P < 0.001